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<p style="text-align: justify;">Previous name: Chromatin Shearing Optimization Kit - High SDS (True Micro ChIP kit)</p>
<p style="text-align: justify;">A high quality chromatin preparation is very complex and requires a lot of optimization. Chromatin EasyShear Kit – High SDS is an optimized solution for efficient chromatin preparation prior to ChIP. The protocol, buffers composition, SDS concentration (1%) is optimized for the preparation of chromatin prior to ChIP on low amount of starting material<b> </b>and it is compatible with Diagenode's <a href="https://www.diagenode.com/en/p/true-microchip-kit-x16-16-rxns">True MicroChIP-seq kit</a> and <a href="https://www.diagenode.com/en/p/uchipmentation-for-histones-24-rxns">µChIPmentation Kit for Histones</a>. The kit has been validated with the Bioruptor ultrasonicator for efficient chromatin shearing, leading to chromatin fragments<span> </span><strong>suitable for ChIP</strong><span> </span>with the preserved<span> </span><strong>epitopes</strong>.</p>
<p style="text-align: justify;">Check all <a href="https://www.diagenode.com/en/categories/chromatin-shearing">Chromatin EasyShear Kits</a>.</p>
<p style="text-align: justify;">Guide for the optimal chromatin preparation using Chromatin EasyShear Kits – <a href="https://www.diagenode.com/en/pages/chromatin-prep-easyshear-kit-guide">Read more</a></p>',
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<p style="text-align: justify;">Previous name: Chromatin Shearing Optimization Kit - High SDS (True Micro ChIP kit)</p>
<p style="text-align: justify;">A high quality chromatin preparation is very complex and requires a lot of optimization. Chromatin EasyShear Kit – High SDS is an optimized solution for efficient chromatin preparation prior to ChIP. The protocol, buffers composition, SDS concentration (1%) is optimized for the preparation of chromatin prior to ChIP on low amount of starting material<b> </b>and it is compatible with Diagenode's <a href="https://www.diagenode.com/en/p/true-microchip-kit-x16-16-rxns">True MicroChIP-seq kit</a> and <a href="https://www.diagenode.com/en/p/uchipmentation-for-histones-24-rxns">µChIPmentation Kit for Histones</a>. The kit has been validated with the Bioruptor ultrasonicator for efficient chromatin shearing, leading to chromatin fragments<span> </span><strong>suitable for ChIP</strong><span> </span>with the preserved<span> </span><strong>epitopes</strong>.</p>
<p style="text-align: justify;">Check all <a href="https://www.diagenode.com/en/categories/chromatin-shearing">Chromatin EasyShear Kits</a>.</p>
<p style="text-align: justify;">Guide for the optimal chromatin preparation using Chromatin EasyShear Kits – <a href="https://www.diagenode.com/en/pages/chromatin-prep-easyshear-kit-guide">Read more</a></p>',
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<p>The <b>True </b><b>MicroChIP-seq</b><b> kit </b>provides a robust ChIP protocol suitable for the investigation of histone modifications within chromatin from as few as <b>10 000 cells</b>, including <b>FACS sorted cells</b>. The kit can be used for chromatin preparation for downstream ChIP-qPCR or ChIP-seq analysis. The <b>complete kit</b> contains everything you need for start-to-finish ChIP including all validated buffers and reagents for chromatin shearing, immunoprecipitation and DNA purification for exceptional <strong>ChIP-qPCR</strong> or <strong>ChIP-seq</strong> results. In addition, positive control antibodies and negative control PCR primers are included for your convenience and assurance of result sensitivity and specificity.</p>
<p>The True MicroChIP-seq kit offers unique benefits:</p>
<ul>
<li>An <b>optimized chromatin preparation </b>protocol compatible with low number of cells (<b>10.000</b>) in combination with the Bioruptor™ shearing device</li>
<li>Most <b>complete kit </b>available (covers all steps and includes control antibodies and primers)</li>
<li><b>Magnetic beads </b>make ChIP easy, fast, and more reproducible</li>
<li>MicroChIP DiaPure columns (included in the kit) enable the <b>maximum recovery </b>of immunoprecipitation DNA suitable for any downstream application</li>
<li><b>Excellent </b><b>ChIP</b><b>-seq </b>result when combined with <a href="https://www.diagenode.com/en/categories/library-preparation-for-ChIP-seq">MicroPlex</a><a href="https://www.diagenode.com/en/categories/library-preparation-for-ChIP-seq"> Library Preparation kit </a>adapted for low input</li>
</ul>
<p>For fast ChIP-seq on low input – check out Diagenode’s <a href="https://www.diagenode.com/en/p/uchipmentation-for-histones-24-rxns">µ</a><a href="https://www.diagenode.com/en/p/uchipmentation-for-histones-24-rxns">ChIPmentation</a><a href="https://www.diagenode.com/en/p/uchipmentation-for-histones-24-rxns"> for histones</a>.</p>
<p><sub>The True MicroChIP-seq kit, Cat. No. C01010132 is an upgraded version of the kit True MicroChIP, Cat. No. C01010130, with the new validated protocols (e.g. FACS sorted cells) and MicroChIP DiaPure columns included in the kit.</sub></p>',
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<li><b>Revolutionary:</b> Only 10,000 cells needed for complete ChIP-seq procedure</li>
<li><b>Validated on</b> studies for histone marks</li>
<li><b>Automated protocol </b>for the IP-Star<sup>®</sup> Compact Automated Platform available</li>
</ul>
<p></p>
<p>The True MicroChIP-seq kit protocol has been optimized for the use of 10,000 - 100,000 cells per immunoprecipitation reaction. Regarding chromatin immunoprecipitation, three protocol variants have been optimized:<br />starting with a batch, starting with an individual sample and starting with the FACS-sorted cells.</p>
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<div>
<h3>High efficiency ChIP on 10,000 cells</h3>
<div class="large-10 small-12 medium-10 large-centered medium-centered small-centered columns"><img src="https://www.diagenode.com/img/product/kits/true-micro-chip-histone-results.png" width="800px" /></div>
<div class="large-10 small-12 medium-10 large-centered medium-centered small-centered columns"><center>
<p><small><strong>Figure 1. </strong>ChIP efficiency on 10,000 cells. ChIP was performed on human Hela cells using the Diagenode antibodies <a href="https://www.diagenode.com/en/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">H3K4me3</a> (Cat. No. C15410003), <a href="https://www.diagenode.com/en/p/h3k27ac-polyclonal-antibody-classic-50-mg-42-ml">H3K27ac</a> (C15410174), <a href="https://www.diagenode.com/en/p/h3k9me3-polyclonal-antibody-classic-50-ug">H3K9me3</a> (C15410056) and <a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-classic-50-mg-34-ml">H3K27me3</a> (C15410069). Sheared chromatin from 10,000 cells and 0.1 µg (H3K27ac), 0.25 µg (H3K4me3 and H3K27me3) or 0.5 µg (H3K9me3) of the antibody were used per IP. Corresponding amount of IgG was used as control. Quantitative PCR was performed with primers for corresponding positive and negative loci. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<div>
<h3>True MicroChIP-seq protocol in a combination with MicroPlex library preparation kit results in reliable and accurate sequencing data</h3>
<div class="large-10 small-12 medium-10 large-centered medium-centered small-centered columns"><img src="https://www.diagenode.com/img/product/kits/fig2-truemicro.jpg" alt="True MicroChip results" width="800px" /></div>
<div class="large-10 small-12 medium-10 large-centered medium-centered small-centered columns"><center>
<p><small><strong>Figure 2.</strong> Integrative genomics viewer (IGV) visualization of ChIP-seq experiments using 50.000 of K562 cells. ChIP has been performed accordingly to True MicroChIP protocol followed by the library preparation using MicroPlex Library Preparation Kit (C05010001). The above figure shows the peaks from ChIP-seq experiments using the following antibodies: H3K4me1 (C15410194), H3K9/14ac (C15410200), H3K27ac (C15410196) and H3K36me3 (C15410192).</small></p>
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<h3>Successful chromatin profiling from 10.000 of FACS-sorted cells</h3>
<div class="large-10 small-12 medium-10 large-centered medium-centered small-centered columns"><img src="https://www.diagenode.com/img/product/kits/fig3ab-truemicro.jpg" alt="small non coding RNA" width="800px" /></div>
<div class="large-10 small-12 medium-10 large-centered medium-centered small-centered columns"><center>
<p><small><strong>Figure 3.</strong> (A) Integrative genomics viewer (IGV) visualization of ChIP-seq experiments and heatmap 3kb upstream and downstream of the TSS (B) for H3K4me3. ChIP has been performed using 10.000 of FACS-sorted cells (K562) and H3K4me3 antibody (C15410003) accordingly to True MicroChIP protocol followed by the library preparation using MicroPlex Library Preparation Kit (C05010001). Data were compared to ENCODE standards.</small></p>
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'label2' => 'Additional solutions compatible with the True MicroChIP-seq Kit',
'info2' => '<p><span style="font-weight: 400;">The <a href="https://www.diagenode.com/en/p/chromatin-shearing-optimization-kit-high-sds-100-million-cells">Chromatin EasyShear Kit – High SDS</a></span><span style="font-weight: 400;"> Recommended for the optimizing chromatin shearing.</span></p>
<p><a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies"><span style="font-weight: 400;">ChIP-seq grade antibodies</span></a><span style="font-weight: 400;"> for high yields, specificity, and sensitivity.</span></p>
<p><span style="font-weight: 400;">Check the list of available </span><a href="https://www.diagenode.com/en/categories/primer-pairs"><span style="font-weight: 400;">primer pairs</span></a><span style="font-weight: 400;"> designed for high specificity to specific genomic regions.</span></p>
<p><span style="font-weight: 400;">For library preparation of immunoprecipitated samples we recommend to use the </span><b> </b><a href="https://www.diagenode.com/en/categories/library-preparation-for-ChIP-seq"><span style="font-weight: 400;">MicroPlex Library Preparation Kit</span></a><span style="font-weight: 400;"> - validated for library preparation from picogram inputs.</span></p>
<p><span style="font-weight: 400;">For IP-Star Automation users, check out the </span><a href="https://www.diagenode.com/en/p/auto-true-microchip-kit-16-rxns"><span style="font-weight: 400;">automated version</span></a><span style="font-weight: 400;"> of this kit.</span></p>
<p><span style="font-weight: 400;">Application note: </span><a href="https://www.diagenode.com/files/application_notes/Diagenode_AATI_Joint.pdf"><span style="font-weight: 400;">Best Workflow Practices for ChIP-seq Analysis with Small Samples</span></a></p>
<p></p>',
'label3' => 'Species, cell lines, tissues tested',
'info3' => '<p>The True MicroChIP-seq kit is compatible with a broad variety of cell lines, tissues and species - some examples are shown below. Other species / cell lines / tissues can be used with this kit.</p>
<p><strong>Cell lines:</strong></p>
<p>Bovine: blastocysts,<br />Drosophila: embryos, salivary glands<br />Human: EndoC-ẞH1 cells, HeLa cells, PBMC, urothelial cells<br />Mouse: adipocytes, B cells, blastocysts, pre-B cells, BMDM cells, chondrocytes, embryonic stem cells, KH2 cells, LSK cells, macrophages, MEP cells, microglia, NK cells, oocytes, pancreatic cells, P19Cl6 cells, RPE cells,</p>
<p>Other cell lines / species: compatible, not tested</p>
<p><strong>Tissues:</strong></p>
<p>Horse: adipose tissue</p>
<p>Mice: intestine tissue</p>
<p>Other tissues: not tested</p>',
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'description' => '<p>The Auto True MicroChIP kit in combination with the <a href="../p/microplex-library-preparation-kit-v2-x12-12-indices-12-rxns">MicroPlex Library Preparation™ kit</a> allows for performing ChIP-seq on as few as 10,000 cells. This microChIP-seq assay has been validated with the new generation <a href="../p/sx-8g-ip-star-compact-automated-system-1-unit">IP-Star<sup>®</sup> Compact Automated Workstation</a>. Furthermore Diagenode's high quality ChIP-seq grade antibodies have been used.</p>
<div class="row">
<div class="small-12 medium-5 large-6 columns"><small style="color: #666;">Video article</small>
<div class="flex-video"><iframe width="420" height="315" style="float: left; margin-right: 20px;" src="//www.youtube.com/embed/JZC9ARbLh_U?rel=0" frameborder="0" allowfullscreen="allowfullscreen"></iframe></div>
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<h3>Automating ChIP-seq Experiments to Generate Epigenetic Profiles on 10,000 HeLa Cells</h3>
<p><a href="https://www.youtube.com/watch?v=JZC9ARbLh_U"><img src="https://www.diagenode.com/img/jove_logo.png" alt="Jove logo" border="0" width="100" height="58" /></a></p>
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<li><strong>Revolutionary:</strong> Only 10.000 cells needed for complete ChIP-seq procedure</li>
<li><strong>Reliable and accurate</strong> sequencing library preparation from just picogram inputs</li>
<li><strong>Validated on</strong> studies for histone marks and with the IP-Star<sup>®</sup> Compact Automated Platform</li>
</ul>
<p><img src="https://www.diagenode.com/img/product/kits/true-micro-chip-histone-results.png" alt="ChIP on 10,000 cells" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><strong>High efficiency ChIP on 10,000 cells</strong><br />ChIP efficiency on 10 000 cells. ChIP was performed on human Hela cells using the Diagenode antibodies H3K4me3 (Cat. No. pAb-003-050), H3K27ac (pAb-174-050), H3K9me3 (pAb-056-050) and H3K27me3 (pAb-069-050). Sheared chromatin from 10 000 cells and 0.1 µg (H3K27ac), 0.25 µg (H3K4me3 and H3K27me3) or 0.5 µg (H3K9me3) of the antibody were used per IP. Corresponding amount of IgG was used as control. Quantitative PCR was performed with primers for corresponding positive and negative loci. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</p>
<p><img src="https://www.diagenode.com/img/product/kits/true-micro-chip-reliable-A.png" alt="ChIP amplification" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/kits/true-micro-chip-reliable-B.png" alt="Match between True MicroChIP peaks and the reference dataset" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><strong>Reliable detection of enrichments in ChIP-seq</strong><br />A: ChIP has been peformed with H3K4me3 antibody, amplification of 17 pg of DNA ChIP'd from 10.000 cells and amplification of 35 pg of DNA ChIP'd from 100.000 cells (control experiment). The IP'd DNA was amplified and transformed into a sequencing-ready preparation for the Illumina plateform with the MicroPlex Library Preparation kit. The library was then analysed on an Illumina<sup>®</sup> Genome Analyzer. Cluster generation and sequencing were performed according to the manufacturer's instructions.<br />B: We observed a perfect match between the top 40% of True MicroChIP peaks and the reference dataset. Based on the NIH Encode project criterion, ChIP-seq results are considered reproducible between an original and reproduced dataset if the top 40% of peaks have at least an 80% overlap ratio with the compared dataset.</p>
<p><img src="https://www.diagenode.com/img/product/kits/true-micro-chip-result.png" alt="Cell lysis" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><strong>Hela cells were fixed with 1% formaldehyde (for 10 minutes at RT)</strong><br /> Cell lysis was performed using the Lysis Buffer tL1 of the Diagenode True MicroChIP kit. Samples corresponding to 10 000 cells are sheared during 5 rounds of 5 cycles of 30 seconds “ON” / 30 seconds “OFF” with the Bioruptor<sup>®</sup> Plus combined with the Bioruptor<sup>®</sup> Water cooler (Cat No. BioAcc-cool) at HIGH power setting (position H). For optimal results, samples are vortexed before and after performing 5 sonication cycles, followed by a short centrifugation at 4°C. 10 μl of DNA (equivalent to 60 000 cells) are analysed on a 1.5% agarose gel.</p>',
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'meta_title' => 'Auto True MicroChIP kit',
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'name' => 'µChIPmentation Kit for Histones',
'description' => '<p><a href="https://www.diagenode.com/files/products/kits/microchipmentation-for-histones.pdf"><img src="https://www.diagenode.com/img/buttons/bt-manual.png" /></a></p>
<p>The Diagenode <strong>µChIPmentation Kit for Histones</strong> is optimized to perform ChIP-seq on as little as <strong>10.000 cells</strong> from cell fixation to purified libraries. To <strong>reduce DNA lost</strong> the number of sample transfer from tube to tube has been limited: during whole workflow - from cell fixation to library prurification - only 3 tubes per sample are used. Reduced number of steps, reduced number of sample transfer and <a href="https://www.diagenode.com/en/categories/chromatin-ip-chipmentation" target="_blank">ChIPmentation technology</a> itself enable for <strong>efficient and robust ChIP-seq</strong> on <strong>limited amount of sample</strong>. The kit µChIPmentation for Histones includes all reagents for chromatin preparation, chromatin immunoprecipitation and library preparation using tagmentation. The primer indexes for multiplexing are not included in the kit and have to be purchase separately - <a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set1">read more</a>.</p>
<div class="extra-spaced">
<h3>Benefits of the µChIPmentation system for histone ChIP-seq</h3>
<ul>
<li><strong>Easier</strong> and <strong>faster</strong> than classical ChIP-seq</li>
<li>Optimized for <strong>low input samples</strong>: as little as 10,000 cells</li>
<li>Protocol optimized for <strong>FACS-sorted cells</strong></li>
<li>Validated for various <strong>histone marks</strong></li>
<li><strong>High quality</strong> sequencing data</li>
</ul>
</div>',
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'info1' => '<p>ChIPmentation is based on tagmentation that allows library preparation to be integrated during the ChIP itself using transposase and sequencing-compatible adaptors. Our new improved µChIPmentation protocol combines 3 features for guaranteeing high quality sequencing data on small sample inputs 1) optimized chromatin shearing preparation protocol, 2) reduced number of steps, 3) reduced number of sample transfer from tube to tube - only 3 tubes per sample for the whole process, from cell fixation to purified libraries.</p>
<ul class="accordion" data-accordion="" style="margin-left: 0;">
<li class="accordion-navigation"><a style="background-color: white;" href="#workflow"><i class="fa fa-caret-right"></i> Workflow of µChIPmentation for Histones - Read more</a>
<div id="workflow" class="content">
<div class="extra-spaced"><center><img src="https://www.diagenode.com/img/product/kits/workflow-micro-chipmentation.png" /></center></div>
</div>
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<div class="row extra-spaced">
<div class="small-12 medium-12 large-12 columns"><strong>A.</strong><img src="https://www.diagenode.com/img/product/kits/micro-chipmentation-fig1A.png" /></div>
</div>
<div class="row extra-spaced">
<div class="small-12 medium-5 large-5 columns" columns=""><strong>B.</strong><img src="https://www.diagenode.com/img/product/kits/micro-chipmentation-fig1B.png" /></div>
<div class="small-12 medium-7 large-7 columns">
<p style="text-align: left;"><strong>Figure 1. Comparison between ChIPmentation and µChIPmentation </strong><br />ChIPmentation: chromatin preparation has been performed on 7 M K562 cells using the ChIPmentation Kit for Histones (Cat. No. C01011000) and diluted chromatin from 500.000 cells was used for the immunoprecipitation. µChIPmentation: chromatin preparation and immunoprecipitation have been performed on 10.000 K562 cells using the µChIPmentation Kit for Histones (Cat. No. C01011011) and 24 SI for Tagmented libraries (Cat. No. C010111032). The Diagenode antibody targeting H3K27me3 (Cat. No. C15410195) was used.<br /> A. Distribution of the ChIPmentation and µChIPmentation readsets in a representative region of the genome (in duplicates). B. Comparison of the top 40% peaks from µChIPmentation (10.000 cells) with dataset generated with ChIPmentation (500.000 cells).</p>
</div>
</div>
<div class="extra-spaced">
<div class="row">
<div class="small-12 medium-12 large-12 columns"><img src="https://www.diagenode.com/img/product/kits/micro-chipmentation-fig2.png" /></div>
</div>
<div class="row">
<div class="small-12 medium-12 large-12 columns">
<p><strong>Figure 2. Sequencing profiles of µChIPmentation libraries </strong><br />Chromatin preparation and immunoprecipitation have been performed on 10.000 cells using the µChIPmentation Kit for Histones (Cat. No. C01011011) and 24 SI for Tagmented libraries (Cat. No. C010111032) using K562 cells. The Diagenode antibodies targeting H3K4me3 (Cat. No. C15410003), H3K27ac (Cat. No. C15410196), H3K27me3 (Cat. No. C15410195) and H3K9me3 (Cat. No. C15410193) have been used.</p>
</div>
</div>
</div>
<div class="extra-spaced">
<div class="row">
<div class="small-12 medium-12 large-12 columns"><img src="https://www.diagenode.com/img/product/kits/micro-chipmentation-fig3.png" /></div>
</div>
<div class="row">
<div class="small-12 medium-12 large-12 columns">
<p><strong>Figure 3. Integrative genomics viewer (IGV) visualization of from ChIP-seq experiments using H3K27me3 antibody (Diagenode, Cat. No. <span>C15410195</span>) and 10.000 cells of K562 cells per immunoprecipitation.<br /></strong>Cells were FACS-sorted and ChIP has been performed accordingly to µChIPmentation protocol. Batch chromatin preparation followed by immunoprecipitation has been used per comparison as indicated.</p>
</div>
</div>
</div>',
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'info2' => '<p><a href="https://www.diagenode.com/en/p/chromatin-shearing-optimization-kit-high-sds-100-million-cells">Chromatin EasyShear Kit - High SDS</a> optimizes chromatin shearing, a critical step for ChIP.</p>
<p><a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies" target="_blank">ChIP-seq grade anti-histone antibodies</a> provide high yields with excellent specificity and sensitivity.</p>
<p>For fast and efficient isolation of magnetic beads we recommend the magnetic racks <a href="https://www.diagenode.com/en/p/diamag02-magnetic-rack-1-unit">DiaMag0.2</a>.</p>
<p>Primer indexes for multiplexing:</p>
<ul class="no-bullet">
<li><a href="https://www.diagenode.com/en/p/24-si-for-tagmented-libraries" target="_blank">24 SI for Tagmented libraries Cat. No. C01011032</a></li>
<li><a href="https://www.diagenode.com/en/p/8-si-for-tagmented-libraries" target="_blank">8 SI for Tagmented libraries Cat. No. C01011033</a></li>
<li><em></em><a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set1">24 UDI for Tagmented libraries - Set I, Cat. No. C0101134</a></li>
<li><a href="https://www.diagenode.com/en/p/8-unique-dual-indexes-for-tagmented-libraries">8 UDI for Tagmented libraries, Cat. No. C0101135</a></li>
<li><a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set2">24 UDI for Tagmented libraries - Set II, Cat. No. C0101136</a></li>
<li><a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set3">24 UDI for Tagmented libraries - Set III, Cat. No. C0101137</a></li>
<li></li>
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<div class="small-12 medium-12 large-12 columns">The most important steps for a successful ChIP include both cell fixation and lysis, and chromatin shearing. Researchers often overlook the critical nature of both of these steps. Eliminating inconsistencies in the shearing step, <strong>Diagenode's Bioruptor</strong><sup>®</sup> uses state-of-the-art ultrasound <strong>ACT</strong> (<strong>A</strong>daptive <strong>C</strong>avitation <strong>T</strong>echnology) to efficiently shear chromatin. ACT enables the highest chromatin quality for high IP efficiency and sensitivity for ChIP experiments with gentle yet highly effective shearing forces. Additionally, the Bioruptor<sup>®</sup> provides a precisely controlled temperature environment that preserves chromatin from heat degradation such that protein-DNA complexes are well-preserved for sensitive, unbiased, and accurate ChIP.<br /><br /> <strong>Diagenode's Bioruptor</strong><sup>®</sup> is the instrument of choice for chromatin shearing used for a number of downstream applications such as qPCR and ChIP-seq that require optimally sheared, unbiased chromatin.</div>
<div class="small-12 medium-12 large-12 columns text-center"><br /><img src="https://www.diagenode.com/img/applications/pico_dna_shearing_fig2.png" /></div>
<div class="small-10 medium-10 large-10 columns end small-offset-1"><small> <br /><strong>Panel A, 10 µl volume:</strong> Chromatin samples are sheared for 10, 20 and 30 cycles of 30 sec ON/30 sec OFF with the Bioruptor® Pico using 0.1 ml Bioruptor® Microtubes (Cat. No. B01200041). <strong>Panel B, 100 µl volume:</strong> Chromatin samples are sheared for 10 cycles of 30 sec ON/30 sec OFF with the Bioruptor® Pico using 0.65 ml Bioruptor® Microtubes (Cat. No. WA-005-0500). <strong>Panel C, 300 µl volume:</strong> Chromatin samples are sheared for 5, 10 and 15 cycles of 30 sec ON/30 sec OFF with the Bioruptor® Pico using using 1.5 ml Bioruptor microtubes (Cat. No. C30010016). Prior to de-crosslinking, samples are treated with RNase cocktail mixture at 37°C during 1 hour. The sheared chromatin is then de-crosslinked overnight and phenol/chloroform purified as described in the kit manual. 10 µl of DNA (equivalent of 500, 000 cells) are analyzed on a 2% agarose gel (MW corresponds to the 100 bp DNA molecular weight marker).</small></div>
<div class="small-12 medium-12 large-12 columns"><br /><br /></div>
<div class="small-12 medium-12 large-12 columns">
<p>It is important to establish optimal conditions to shear crosslinked chromatin to get the correct fragment sizes needed for ChIP. Usually this process requires both optimizing sonication conditions as well as optimizing SDS concentration, which is laborious. With the Chromatin Shearing Optimization Kits, optimization is fast and easy - we provide optimization reagents with varying concentrations of SDS. Moreover, our Chromatin Shearing Optimization Kits can be used for the optimization of chromatin preparation with our kits for ChIP.</p>
</div>
<div class="small-12 medium-12 large-12 columns">
<div class="page" title="Page 7">
<table>
<tbody>
<tr valign="middle">
<td></td>
<td style="text-align: center;"><strong><a href="https://www.diagenode.com/p/chromatin-shearing-optimization-kit-low-sds-100-million-cells">Chromatin Shearing Kit Low SDS (for Histone)</a></strong></td>
<td style="text-align: center;"><strong><a href="https://www.diagenode.com/p/chromatin-shearing-optimization-kit-low-sds-for-tfs-25-rxns">Chromatin Shearing Kit Low SDS (for TF)</a></strong></td>
<td style="text-align: center;"><strong><a href="https://www.diagenode.com/p/chromatin-shearing-optimization-kit-high-sds-100-million-cells">Chromatin Shearing Kit High SDS</a></strong></td>
<td style="text-align: center;"><strong><a href="https://www.diagenode.com/p/chromatin-shearing-optimization-kit-medium-sds-100-million-cells">Chromatin Shearing Kit (for Plant)</a></strong></td>
</tr>
<tr valign="middle" style="background-color: #fff;">
<td>
<p><strong>SDS concentration</strong></p>
</td>
<td style="text-align: center;">
<p>< 0.1%</p>
</td>
<td style="text-align: center;">
<p>0.2%</p>
</td>
<td style="text-align: center;">
<p>1%</p>
</td>
<td style="text-align: center;">
<p>0.5%</p>
</td>
</tr>
<tr valign="middle" style="background-color: #fff;">
<td>
<p><strong>Nuclei isolation</strong></p>
</td>
<td style="text-align: center;">
<p>Yes</p>
</td>
<td style="text-align: center;">
<p>Yes</p>
</td>
<td style="text-align: center;">
<p>No</p>
</td>
<td style="text-align: center;">
<p>Yes</p>
</td>
</tr>
<tr valign="middle" style="background-color: #fff;">
<td>
<p><strong>Allows for shearing of... cells/tissue</strong></p>
</td>
<td style="text-align: center;">
<p>100 million cells</p>
</td>
<td style="text-align: center;">
<p>100 million cells</p>
</td>
<td style="text-align: center;">
<p>100 million cells</p>
</td>
<td style="text-align: center;">
<p>up to 25 g of tissue</p>
</td>
</tr>
<tr valign="middle" style="background-color: #fff;">
<td>
<p><strong>Corresponding to shearing buffers from</strong></p>
</td>
<td style="text-align: center;">
<p><a href="https://www.diagenode.com/en/p/ideal-chip-seq-kit-x24-24-rxns">iDeal ChIP-seq kit for Histones</a></p>
</td>
<td style="text-align: center;">
<p><a href="https://www.diagenode.com/en/p/ideal-chip-seq-kit-for-transcription-factors-x24-24-rxns">iDeal ChIP-seq Kit for Transcription Factors</a></p>
<p><a href="https://www.diagenode.com/en/p/ideal-chip-qpcr-kit">iDeal ChIP qPCR kit</a></p>
</td>
<td style="text-align: center;">
<p><a href="https://www.diagenode.com/en/p/true-microchip-kit-x16-16-rxns">True MicroChIP kit</a></p>
</td>
<td style="text-align: center;">
<p><a href="https://www.diagenode.com/en/p/universal-plant-chip-seq-kit-x24-24-rxns">Universal Plant ChIP-seq kit</a></p>
</td>
</tr>
</tbody>
</table>
<p><em><span style="font-weight: 400;">Table comes from our </span><a href="https://www.diagenode.com/protocols/bioruptor-pico-chromatin-preparation-guide"><span style="font-weight: 400;">Guide for successful chromatin preparation using the Bioruptor® Pico</span></a></em></p>
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'description' => '<p>Optimal detergent concentration is required for best results with chromatin shearing. Diagenode offers four kits with different SDS concentrations that have been pre-optimized for your specific workflow requirements. Our Chromatin EasyShear Kits (previous name: Chromatin Shearing Optimization Kits) together with the Bioruptor combine efficient cell lysis and chromatin shearing leading to consistent results.</p>
<p>The Chromatin EasyShear Kits are recommended for:</p>
<ul>
<li>The optimization of the chromatin shearing and/or chromatin preparation prior to any ChIP protocol</li>
<li>The optimization of the chromatin shearing of a new cell line/new sample type prior to ChIP using Diagenode’s ChIP kits</li>
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<p>Check all <a href="https://www.diagenode.com/en/categories/chromatin-shearing">Chromatin EasyShear Kits</a>.</p>
<p>Guide for the optimal chromatin preparation using Chromatin EasyShear Kits – <a href="https://www.diagenode.com/en/pages/chromatin-prep-easyshear-kit-guide">Read more</a></p>
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<p><span>The first critical step of a successful Chromatin immunoprecipitation (ChIP) experiment is the preparation of sheared chromatin. We therefore suggest to use one of our optimized shearing ChIP kits.</span></p>
<span>• Chromatin EasyShear Kit - Ultra Low SDS <br />• Chromatin <span>EasyShear Kit</span> <span>-</span> Low SDS <br />• Chromatin EasyShear Kit - High SDS </span></div>
<div class="column"><span>• Chromatin EasyShear Kit for Plant</span><span></span>
<p><span></span></p>
<p><span>Our Chromatin EasyShear kits are used in combination with the Bioruptor</span><span>® </span><span>in order to ensure a highly reproducible chromatin shearing and make sure you obtain the right fragment size needed for your experiment. Establish optimal conditions required for shearing cross-linked chromatin (protein-DNA) is usually laborious; the protocol of the Chromatin EasyShear kits is fast, easy-to-use and optimized to get the best results possible. </span></p>
<p><span>Learn more about Diagenode <a href="https://www.diagenode.com/en/categories/chromatin-shearing">Chromatin EasyShear Kits</a>.</span></p>
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<p>The Diagenode <strong>µChIPmentation Kit for Histones</strong> is optimized to perform ChIP-seq on as little as <strong>10.000 cells</strong> from cell fixation to purified libraries. To <strong>reduce DNA lost</strong> the number of sample transfer from tube to tube has been limited: during whole workflow - from cell fixation to library prurification - only 3 tubes per sample are used. Reduced number of steps, reduced number of sample transfer and <a href="https://www.diagenode.com/en/categories/chromatin-ip-chipmentation" target="_blank">ChIPmentation technology</a> itself enable for <strong>efficient and robust ChIP-seq</strong> on <strong>limited amount of sample</strong>. The kit µChIPmentation for Histones includes all reagents for chromatin preparation, chromatin immunoprecipitation and library preparation using tagmentation. The primer indexes for multiplexing are not included in the kit and have to be purchase separately - <a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set1">read more</a>.</p>
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<ul class="accordion" data-accordion="" style="margin-left: 0;">
<li class="accordion-navigation"><a style="background-color: white;" href="#workflow"><i class="fa fa-caret-right"></i> Workflow of µChIPmentation for Histones - Read more</a>
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<p style="text-align: left;"><strong>Figure 1. Comparison between ChIPmentation and µChIPmentation </strong><br />ChIPmentation: chromatin preparation has been performed on 7 M K562 cells using the ChIPmentation Kit for Histones (Cat. No. C01011000) and diluted chromatin from 500.000 cells was used for the immunoprecipitation. µChIPmentation: chromatin preparation and immunoprecipitation have been performed on 10.000 K562 cells using the µChIPmentation Kit for Histones (Cat. No. C01011011) and 24 SI for Tagmented libraries (Cat. No. C010111032). The Diagenode antibody targeting H3K27me3 (Cat. No. C15410195) was used.<br /> A. Distribution of the ChIPmentation and µChIPmentation readsets in a representative region of the genome (in duplicates). B. Comparison of the top 40% peaks from µChIPmentation (10.000 cells) with dataset generated with ChIPmentation (500.000 cells).</p>
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<p><strong>Figure 2. Sequencing profiles of µChIPmentation libraries </strong><br />Chromatin preparation and immunoprecipitation have been performed on 10.000 cells using the µChIPmentation Kit for Histones (Cat. No. C01011011) and 24 SI for Tagmented libraries (Cat. No. C010111032) using K562 cells. The Diagenode antibodies targeting H3K4me3 (Cat. No. C15410003), H3K27ac (Cat. No. C15410196), H3K27me3 (Cat. No. C15410195) and H3K9me3 (Cat. No. C15410193) have been used.</p>
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<p><strong>Figure 3. Integrative genomics viewer (IGV) visualization of from ChIP-seq experiments using H3K27me3 antibody (Diagenode, Cat. No. <span>C15410195</span>) and 10.000 cells of K562 cells per immunoprecipitation.<br /></strong>Cells were FACS-sorted and ChIP has been performed accordingly to µChIPmentation protocol. Batch chromatin preparation followed by immunoprecipitation has been used per comparison as indicated.</p>
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<p><a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies" target="_blank">ChIP-seq grade anti-histone antibodies</a> provide high yields with excellent specificity and sensitivity.</p>
<p>For fast and efficient isolation of magnetic beads we recommend the magnetic racks <a href="https://www.diagenode.com/en/p/diamag02-magnetic-rack-1-unit">DiaMag0.2</a>.</p>
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<li><a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set3">24 UDI for Tagmented libraries - Set III, Cat. No. C0101137</a></li>
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<p><span>The first critical step of a successful Chromatin immunoprecipitation (ChIP) experiment is the preparation of sheared chromatin. We therefore suggest to use one of our optimized shearing ChIP kits.</span></p>
<span>• Chromatin EasyShear Kit - Ultra Low SDS <br />• Chromatin <span>EasyShear Kit</span> <span>-</span> Low SDS <br />• Chromatin EasyShear Kit - High SDS </span></div>
<div class="column"><span>• Chromatin EasyShear Kit for Plant</span><span></span>
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<p><span>Our Chromatin EasyShear kits are used in combination with the Bioruptor</span><span>® </span><span>in order to ensure a highly reproducible chromatin shearing and make sure you obtain the right fragment size needed for your experiment. Establish optimal conditions required for shearing cross-linked chromatin (protein-DNA) is usually laborious; the protocol of the Chromatin EasyShear kits is fast, easy-to-use and optimized to get the best results possible. </span></p>
<p><span>Learn more about Diagenode <a href="https://www.diagenode.com/en/categories/chromatin-shearing">Chromatin EasyShear Kits</a>.</span></p>
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<p style="text-align: justify;">Previous name: Chromatin Shearing Optimization Kit - High SDS (True Micro ChIP kit)</p>
<p style="text-align: justify;">A high quality chromatin preparation is very complex and requires a lot of optimization. Chromatin EasyShear Kit – High SDS is an optimized solution for efficient chromatin preparation prior to ChIP. The protocol, buffers composition, SDS concentration (1%) is optimized for the preparation of chromatin prior to ChIP on low amount of starting material<b> </b>and it is compatible with Diagenode's <a href="https://www.diagenode.com/en/p/true-microchip-kit-x16-16-rxns">True MicroChIP-seq kit</a> and <a href="https://www.diagenode.com/en/p/uchipmentation-for-histones-24-rxns">µChIPmentation Kit for Histones</a>. The kit has been validated with the Bioruptor ultrasonicator for efficient chromatin shearing, leading to chromatin fragments<span> </span><strong>suitable for ChIP</strong><span> </span>with the preserved<span> </span><strong>epitopes</strong>.</p>
<p style="text-align: justify;">Check all <a href="https://www.diagenode.com/en/categories/chromatin-shearing">Chromatin EasyShear Kits</a>.</p>
<p style="text-align: justify;">Guide for the optimal chromatin preparation using Chromatin EasyShear Kits – <a href="https://www.diagenode.com/en/pages/chromatin-prep-easyshear-kit-guide">Read more</a></p>',
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<p style="text-align: justify;">Previous name: Chromatin Shearing Optimization Kit - High SDS (True Micro ChIP kit)</p>
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<p style="text-align: justify;">Check all <a href="https://www.diagenode.com/en/categories/chromatin-shearing">Chromatin EasyShear Kits</a>.</p>
<p style="text-align: justify;">Guide for the optimal chromatin preparation using Chromatin EasyShear Kits – <a href="https://www.diagenode.com/en/pages/chromatin-prep-easyshear-kit-guide">Read more</a></p>',
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<p>The <b>True </b><b>MicroChIP-seq</b><b> kit </b>provides a robust ChIP protocol suitable for the investigation of histone modifications within chromatin from as few as <b>10 000 cells</b>, including <b>FACS sorted cells</b>. The kit can be used for chromatin preparation for downstream ChIP-qPCR or ChIP-seq analysis. The <b>complete kit</b> contains everything you need for start-to-finish ChIP including all validated buffers and reagents for chromatin shearing, immunoprecipitation and DNA purification for exceptional <strong>ChIP-qPCR</strong> or <strong>ChIP-seq</strong> results. In addition, positive control antibodies and negative control PCR primers are included for your convenience and assurance of result sensitivity and specificity.</p>
<p>The True MicroChIP-seq kit offers unique benefits:</p>
<ul>
<li>An <b>optimized chromatin preparation </b>protocol compatible with low number of cells (<b>10.000</b>) in combination with the Bioruptor™ shearing device</li>
<li>Most <b>complete kit </b>available (covers all steps and includes control antibodies and primers)</li>
<li><b>Magnetic beads </b>make ChIP easy, fast, and more reproducible</li>
<li>MicroChIP DiaPure columns (included in the kit) enable the <b>maximum recovery </b>of immunoprecipitation DNA suitable for any downstream application</li>
<li><b>Excellent </b><b>ChIP</b><b>-seq </b>result when combined with <a href="https://www.diagenode.com/en/categories/library-preparation-for-ChIP-seq">MicroPlex</a><a href="https://www.diagenode.com/en/categories/library-preparation-for-ChIP-seq"> Library Preparation kit </a>adapted for low input</li>
</ul>
<p>For fast ChIP-seq on low input – check out Diagenode’s <a href="https://www.diagenode.com/en/p/uchipmentation-for-histones-24-rxns">µ</a><a href="https://www.diagenode.com/en/p/uchipmentation-for-histones-24-rxns">ChIPmentation</a><a href="https://www.diagenode.com/en/p/uchipmentation-for-histones-24-rxns"> for histones</a>.</p>
<p><sub>The True MicroChIP-seq kit, Cat. No. C01010132 is an upgraded version of the kit True MicroChIP, Cat. No. C01010130, with the new validated protocols (e.g. FACS sorted cells) and MicroChIP DiaPure columns included in the kit.</sub></p>',
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<li><b>Revolutionary:</b> Only 10,000 cells needed for complete ChIP-seq procedure</li>
<li><b>Validated on</b> studies for histone marks</li>
<li><b>Automated protocol </b>for the IP-Star<sup>®</sup> Compact Automated Platform available</li>
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<p></p>
<p>The True MicroChIP-seq kit protocol has been optimized for the use of 10,000 - 100,000 cells per immunoprecipitation reaction. Regarding chromatin immunoprecipitation, three protocol variants have been optimized:<br />starting with a batch, starting with an individual sample and starting with the FACS-sorted cells.</p>
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<div class="large-12 columns truemicro-slider" id="truemicro-slider">
<div>
<h3>High efficiency ChIP on 10,000 cells</h3>
<div class="large-10 small-12 medium-10 large-centered medium-centered small-centered columns"><img src="https://www.diagenode.com/img/product/kits/true-micro-chip-histone-results.png" width="800px" /></div>
<div class="large-10 small-12 medium-10 large-centered medium-centered small-centered columns"><center>
<p><small><strong>Figure 1. </strong>ChIP efficiency on 10,000 cells. ChIP was performed on human Hela cells using the Diagenode antibodies <a href="https://www.diagenode.com/en/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">H3K4me3</a> (Cat. No. C15410003), <a href="https://www.diagenode.com/en/p/h3k27ac-polyclonal-antibody-classic-50-mg-42-ml">H3K27ac</a> (C15410174), <a href="https://www.diagenode.com/en/p/h3k9me3-polyclonal-antibody-classic-50-ug">H3K9me3</a> (C15410056) and <a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-classic-50-mg-34-ml">H3K27me3</a> (C15410069). Sheared chromatin from 10,000 cells and 0.1 µg (H3K27ac), 0.25 µg (H3K4me3 and H3K27me3) or 0.5 µg (H3K9me3) of the antibody were used per IP. Corresponding amount of IgG was used as control. Quantitative PCR was performed with primers for corresponding positive and negative loci. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
</center></div>
</div>
<div>
<h3>True MicroChIP-seq protocol in a combination with MicroPlex library preparation kit results in reliable and accurate sequencing data</h3>
<div class="large-10 small-12 medium-10 large-centered medium-centered small-centered columns"><img src="https://www.diagenode.com/img/product/kits/fig2-truemicro.jpg" alt="True MicroChip results" width="800px" /></div>
<div class="large-10 small-12 medium-10 large-centered medium-centered small-centered columns"><center>
<p><small><strong>Figure 2.</strong> Integrative genomics viewer (IGV) visualization of ChIP-seq experiments using 50.000 of K562 cells. ChIP has been performed accordingly to True MicroChIP protocol followed by the library preparation using MicroPlex Library Preparation Kit (C05010001). The above figure shows the peaks from ChIP-seq experiments using the following antibodies: H3K4me1 (C15410194), H3K9/14ac (C15410200), H3K27ac (C15410196) and H3K36me3 (C15410192).</small></p>
</center></div>
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<div>
<h3>Successful chromatin profiling from 10.000 of FACS-sorted cells</h3>
<div class="large-10 small-12 medium-10 large-centered medium-centered small-centered columns"><img src="https://www.diagenode.com/img/product/kits/fig3ab-truemicro.jpg" alt="small non coding RNA" width="800px" /></div>
<div class="large-10 small-12 medium-10 large-centered medium-centered small-centered columns"><center>
<p><small><strong>Figure 3.</strong> (A) Integrative genomics viewer (IGV) visualization of ChIP-seq experiments and heatmap 3kb upstream and downstream of the TSS (B) for H3K4me3. ChIP has been performed using 10.000 of FACS-sorted cells (K562) and H3K4me3 antibody (C15410003) accordingly to True MicroChIP protocol followed by the library preparation using MicroPlex Library Preparation Kit (C05010001). Data were compared to ENCODE standards.</small></p>
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'label2' => 'Additional solutions compatible with the True MicroChIP-seq Kit',
'info2' => '<p><span style="font-weight: 400;">The <a href="https://www.diagenode.com/en/p/chromatin-shearing-optimization-kit-high-sds-100-million-cells">Chromatin EasyShear Kit – High SDS</a></span><span style="font-weight: 400;"> Recommended for the optimizing chromatin shearing.</span></p>
<p><a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies"><span style="font-weight: 400;">ChIP-seq grade antibodies</span></a><span style="font-weight: 400;"> for high yields, specificity, and sensitivity.</span></p>
<p><span style="font-weight: 400;">Check the list of available </span><a href="https://www.diagenode.com/en/categories/primer-pairs"><span style="font-weight: 400;">primer pairs</span></a><span style="font-weight: 400;"> designed for high specificity to specific genomic regions.</span></p>
<p><span style="font-weight: 400;">For library preparation of immunoprecipitated samples we recommend to use the </span><b> </b><a href="https://www.diagenode.com/en/categories/library-preparation-for-ChIP-seq"><span style="font-weight: 400;">MicroPlex Library Preparation Kit</span></a><span style="font-weight: 400;"> - validated for library preparation from picogram inputs.</span></p>
<p><span style="font-weight: 400;">For IP-Star Automation users, check out the </span><a href="https://www.diagenode.com/en/p/auto-true-microchip-kit-16-rxns"><span style="font-weight: 400;">automated version</span></a><span style="font-weight: 400;"> of this kit.</span></p>
<p><span style="font-weight: 400;">Application note: </span><a href="https://www.diagenode.com/files/application_notes/Diagenode_AATI_Joint.pdf"><span style="font-weight: 400;">Best Workflow Practices for ChIP-seq Analysis with Small Samples</span></a></p>
<p></p>',
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'info3' => '<p>The True MicroChIP-seq kit is compatible with a broad variety of cell lines, tissues and species - some examples are shown below. Other species / cell lines / tissues can be used with this kit.</p>
<p><strong>Cell lines:</strong></p>
<p>Bovine: blastocysts,<br />Drosophila: embryos, salivary glands<br />Human: EndoC-ẞH1 cells, HeLa cells, PBMC, urothelial cells<br />Mouse: adipocytes, B cells, blastocysts, pre-B cells, BMDM cells, chondrocytes, embryonic stem cells, KH2 cells, LSK cells, macrophages, MEP cells, microglia, NK cells, oocytes, pancreatic cells, P19Cl6 cells, RPE cells,</p>
<p>Other cell lines / species: compatible, not tested</p>
<p><strong>Tissues:</strong></p>
<p>Horse: adipose tissue</p>
<p>Mice: intestine tissue</p>
<p>Other tissues: not tested</p>',
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'description' => '<p>The Auto True MicroChIP kit in combination with the <a href="../p/microplex-library-preparation-kit-v2-x12-12-indices-12-rxns">MicroPlex Library Preparation™ kit</a> allows for performing ChIP-seq on as few as 10,000 cells. This microChIP-seq assay has been validated with the new generation <a href="../p/sx-8g-ip-star-compact-automated-system-1-unit">IP-Star<sup>®</sup> Compact Automated Workstation</a>. Furthermore Diagenode's high quality ChIP-seq grade antibodies have been used.</p>
<div class="row">
<div class="small-12 medium-5 large-6 columns"><small style="color: #666;">Video article</small>
<div class="flex-video"><iframe width="420" height="315" style="float: left; margin-right: 20px;" src="//www.youtube.com/embed/JZC9ARbLh_U?rel=0" frameborder="0" allowfullscreen="allowfullscreen"></iframe></div>
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<h3>Automating ChIP-seq Experiments to Generate Epigenetic Profiles on 10,000 HeLa Cells</h3>
<p><a href="https://www.youtube.com/watch?v=JZC9ARbLh_U"><img src="https://www.diagenode.com/img/jove_logo.png" alt="Jove logo" border="0" width="100" height="58" /></a></p>
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<li><strong>Revolutionary:</strong> Only 10.000 cells needed for complete ChIP-seq procedure</li>
<li><strong>Reliable and accurate</strong> sequencing library preparation from just picogram inputs</li>
<li><strong>Validated on</strong> studies for histone marks and with the IP-Star<sup>®</sup> Compact Automated Platform</li>
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<p><img src="https://www.diagenode.com/img/product/kits/true-micro-chip-histone-results.png" alt="ChIP on 10,000 cells" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><strong>High efficiency ChIP on 10,000 cells</strong><br />ChIP efficiency on 10 000 cells. ChIP was performed on human Hela cells using the Diagenode antibodies H3K4me3 (Cat. No. pAb-003-050), H3K27ac (pAb-174-050), H3K9me3 (pAb-056-050) and H3K27me3 (pAb-069-050). Sheared chromatin from 10 000 cells and 0.1 µg (H3K27ac), 0.25 µg (H3K4me3 and H3K27me3) or 0.5 µg (H3K9me3) of the antibody were used per IP. Corresponding amount of IgG was used as control. Quantitative PCR was performed with primers for corresponding positive and negative loci. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</p>
<p><img src="https://www.diagenode.com/img/product/kits/true-micro-chip-reliable-A.png" alt="ChIP amplification" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/kits/true-micro-chip-reliable-B.png" alt="Match between True MicroChIP peaks and the reference dataset" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><strong>Reliable detection of enrichments in ChIP-seq</strong><br />A: ChIP has been peformed with H3K4me3 antibody, amplification of 17 pg of DNA ChIP'd from 10.000 cells and amplification of 35 pg of DNA ChIP'd from 100.000 cells (control experiment). The IP'd DNA was amplified and transformed into a sequencing-ready preparation for the Illumina plateform with the MicroPlex Library Preparation kit. The library was then analysed on an Illumina<sup>®</sup> Genome Analyzer. Cluster generation and sequencing were performed according to the manufacturer's instructions.<br />B: We observed a perfect match between the top 40% of True MicroChIP peaks and the reference dataset. Based on the NIH Encode project criterion, ChIP-seq results are considered reproducible between an original and reproduced dataset if the top 40% of peaks have at least an 80% overlap ratio with the compared dataset.</p>
<p><img src="https://www.diagenode.com/img/product/kits/true-micro-chip-result.png" alt="Cell lysis" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><strong>Hela cells were fixed with 1% formaldehyde (for 10 minutes at RT)</strong><br /> Cell lysis was performed using the Lysis Buffer tL1 of the Diagenode True MicroChIP kit. Samples corresponding to 10 000 cells are sheared during 5 rounds of 5 cycles of 30 seconds “ON” / 30 seconds “OFF” with the Bioruptor<sup>®</sup> Plus combined with the Bioruptor<sup>®</sup> Water cooler (Cat No. BioAcc-cool) at HIGH power setting (position H). For optimal results, samples are vortexed before and after performing 5 sonication cycles, followed by a short centrifugation at 4°C. 10 μl of DNA (equivalent to 60 000 cells) are analysed on a 1.5% agarose gel.</p>',
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'meta_title' => 'Auto True MicroChIP kit',
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'meta_description' => 'Auto True MicroChIP kit',
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'name' => 'µChIPmentation Kit for Histones',
'description' => '<p><a href="https://www.diagenode.com/files/products/kits/microchipmentation-for-histones.pdf"><img src="https://www.diagenode.com/img/buttons/bt-manual.png" /></a></p>
<p>The Diagenode <strong>µChIPmentation Kit for Histones</strong> is optimized to perform ChIP-seq on as little as <strong>10.000 cells</strong> from cell fixation to purified libraries. To <strong>reduce DNA lost</strong> the number of sample transfer from tube to tube has been limited: during whole workflow - from cell fixation to library prurification - only 3 tubes per sample are used. Reduced number of steps, reduced number of sample transfer and <a href="https://www.diagenode.com/en/categories/chromatin-ip-chipmentation" target="_blank">ChIPmentation technology</a> itself enable for <strong>efficient and robust ChIP-seq</strong> on <strong>limited amount of sample</strong>. The kit µChIPmentation for Histones includes all reagents for chromatin preparation, chromatin immunoprecipitation and library preparation using tagmentation. The primer indexes for multiplexing are not included in the kit and have to be purchase separately - <a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set1">read more</a>.</p>
<div class="extra-spaced">
<h3>Benefits of the µChIPmentation system for histone ChIP-seq</h3>
<ul>
<li><strong>Easier</strong> and <strong>faster</strong> than classical ChIP-seq</li>
<li>Optimized for <strong>low input samples</strong>: as little as 10,000 cells</li>
<li>Protocol optimized for <strong>FACS-sorted cells</strong></li>
<li>Validated for various <strong>histone marks</strong></li>
<li><strong>High quality</strong> sequencing data</li>
</ul>
</div>',
'label1' => 'Characteristics',
'info1' => '<p>ChIPmentation is based on tagmentation that allows library preparation to be integrated during the ChIP itself using transposase and sequencing-compatible adaptors. Our new improved µChIPmentation protocol combines 3 features for guaranteeing high quality sequencing data on small sample inputs 1) optimized chromatin shearing preparation protocol, 2) reduced number of steps, 3) reduced number of sample transfer from tube to tube - only 3 tubes per sample for the whole process, from cell fixation to purified libraries.</p>
<ul class="accordion" data-accordion="" style="margin-left: 0;">
<li class="accordion-navigation"><a style="background-color: white;" href="#workflow"><i class="fa fa-caret-right"></i> Workflow of µChIPmentation for Histones - Read more</a>
<div id="workflow" class="content">
<div class="extra-spaced"><center><img src="https://www.diagenode.com/img/product/kits/workflow-micro-chipmentation.png" /></center></div>
</div>
</li>
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<div class="row extra-spaced">
<div class="small-12 medium-12 large-12 columns"><strong>A.</strong><img src="https://www.diagenode.com/img/product/kits/micro-chipmentation-fig1A.png" /></div>
</div>
<div class="row extra-spaced">
<div class="small-12 medium-5 large-5 columns" columns=""><strong>B.</strong><img src="https://www.diagenode.com/img/product/kits/micro-chipmentation-fig1B.png" /></div>
<div class="small-12 medium-7 large-7 columns">
<p style="text-align: left;"><strong>Figure 1. Comparison between ChIPmentation and µChIPmentation </strong><br />ChIPmentation: chromatin preparation has been performed on 7 M K562 cells using the ChIPmentation Kit for Histones (Cat. No. C01011000) and diluted chromatin from 500.000 cells was used for the immunoprecipitation. µChIPmentation: chromatin preparation and immunoprecipitation have been performed on 10.000 K562 cells using the µChIPmentation Kit for Histones (Cat. No. C01011011) and 24 SI for Tagmented libraries (Cat. No. C010111032). The Diagenode antibody targeting H3K27me3 (Cat. No. C15410195) was used.<br /> A. Distribution of the ChIPmentation and µChIPmentation readsets in a representative region of the genome (in duplicates). B. Comparison of the top 40% peaks from µChIPmentation (10.000 cells) with dataset generated with ChIPmentation (500.000 cells).</p>
</div>
</div>
<div class="extra-spaced">
<div class="row">
<div class="small-12 medium-12 large-12 columns"><img src="https://www.diagenode.com/img/product/kits/micro-chipmentation-fig2.png" /></div>
</div>
<div class="row">
<div class="small-12 medium-12 large-12 columns">
<p><strong>Figure 2. Sequencing profiles of µChIPmentation libraries </strong><br />Chromatin preparation and immunoprecipitation have been performed on 10.000 cells using the µChIPmentation Kit for Histones (Cat. No. C01011011) and 24 SI for Tagmented libraries (Cat. No. C010111032) using K562 cells. The Diagenode antibodies targeting H3K4me3 (Cat. No. C15410003), H3K27ac (Cat. No. C15410196), H3K27me3 (Cat. No. C15410195) and H3K9me3 (Cat. No. C15410193) have been used.</p>
</div>
</div>
</div>
<div class="extra-spaced">
<div class="row">
<div class="small-12 medium-12 large-12 columns"><img src="https://www.diagenode.com/img/product/kits/micro-chipmentation-fig3.png" /></div>
</div>
<div class="row">
<div class="small-12 medium-12 large-12 columns">
<p><strong>Figure 3. Integrative genomics viewer (IGV) visualization of from ChIP-seq experiments using H3K27me3 antibody (Diagenode, Cat. No. <span>C15410195</span>) and 10.000 cells of K562 cells per immunoprecipitation.<br /></strong>Cells were FACS-sorted and ChIP has been performed accordingly to µChIPmentation protocol. Batch chromatin preparation followed by immunoprecipitation has been used per comparison as indicated.</p>
</div>
</div>
</div>',
'label2' => 'Additional solutions for µChIPmentation Kit for Histones',
'info2' => '<p><a href="https://www.diagenode.com/en/p/chromatin-shearing-optimization-kit-high-sds-100-million-cells">Chromatin EasyShear Kit - High SDS</a> optimizes chromatin shearing, a critical step for ChIP.</p>
<p><a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies" target="_blank">ChIP-seq grade anti-histone antibodies</a> provide high yields with excellent specificity and sensitivity.</p>
<p>For fast and efficient isolation of magnetic beads we recommend the magnetic racks <a href="https://www.diagenode.com/en/p/diamag02-magnetic-rack-1-unit">DiaMag0.2</a>.</p>
<p>Primer indexes for multiplexing:</p>
<ul class="no-bullet">
<li><a href="https://www.diagenode.com/en/p/24-si-for-tagmented-libraries" target="_blank">24 SI for Tagmented libraries Cat. No. C01011032</a></li>
<li><a href="https://www.diagenode.com/en/p/8-si-for-tagmented-libraries" target="_blank">8 SI for Tagmented libraries Cat. No. C01011033</a></li>
<li><em></em><a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set1">24 UDI for Tagmented libraries - Set I, Cat. No. C0101134</a></li>
<li><a href="https://www.diagenode.com/en/p/8-unique-dual-indexes-for-tagmented-libraries">8 UDI for Tagmented libraries, Cat. No. C0101135</a></li>
<li><a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set2">24 UDI for Tagmented libraries - Set II, Cat. No. C0101136</a></li>
<li><a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set3">24 UDI for Tagmented libraries - Set III, Cat. No. C0101137</a></li>
<li></li>
</ul>',
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<div class="small-12 medium-12 large-12 columns">The most important steps for a successful ChIP include both cell fixation and lysis, and chromatin shearing. Researchers often overlook the critical nature of both of these steps. Eliminating inconsistencies in the shearing step, <strong>Diagenode's Bioruptor</strong><sup>®</sup> uses state-of-the-art ultrasound <strong>ACT</strong> (<strong>A</strong>daptive <strong>C</strong>avitation <strong>T</strong>echnology) to efficiently shear chromatin. ACT enables the highest chromatin quality for high IP efficiency and sensitivity for ChIP experiments with gentle yet highly effective shearing forces. Additionally, the Bioruptor<sup>®</sup> provides a precisely controlled temperature environment that preserves chromatin from heat degradation such that protein-DNA complexes are well-preserved for sensitive, unbiased, and accurate ChIP.<br /><br /> <strong>Diagenode's Bioruptor</strong><sup>®</sup> is the instrument of choice for chromatin shearing used for a number of downstream applications such as qPCR and ChIP-seq that require optimally sheared, unbiased chromatin.</div>
<div class="small-12 medium-12 large-12 columns text-center"><br /><img src="https://www.diagenode.com/img/applications/pico_dna_shearing_fig2.png" /></div>
<div class="small-10 medium-10 large-10 columns end small-offset-1"><small> <br /><strong>Panel A, 10 µl volume:</strong> Chromatin samples are sheared for 10, 20 and 30 cycles of 30 sec ON/30 sec OFF with the Bioruptor® Pico using 0.1 ml Bioruptor® Microtubes (Cat. No. B01200041). <strong>Panel B, 100 µl volume:</strong> Chromatin samples are sheared for 10 cycles of 30 sec ON/30 sec OFF with the Bioruptor® Pico using 0.65 ml Bioruptor® Microtubes (Cat. No. WA-005-0500). <strong>Panel C, 300 µl volume:</strong> Chromatin samples are sheared for 5, 10 and 15 cycles of 30 sec ON/30 sec OFF with the Bioruptor® Pico using using 1.5 ml Bioruptor microtubes (Cat. No. C30010016). Prior to de-crosslinking, samples are treated with RNase cocktail mixture at 37°C during 1 hour. The sheared chromatin is then de-crosslinked overnight and phenol/chloroform purified as described in the kit manual. 10 µl of DNA (equivalent of 500, 000 cells) are analyzed on a 2% agarose gel (MW corresponds to the 100 bp DNA molecular weight marker).</small></div>
<div class="small-12 medium-12 large-12 columns"><br /><br /></div>
<div class="small-12 medium-12 large-12 columns">
<p>It is important to establish optimal conditions to shear crosslinked chromatin to get the correct fragment sizes needed for ChIP. Usually this process requires both optimizing sonication conditions as well as optimizing SDS concentration, which is laborious. With the Chromatin Shearing Optimization Kits, optimization is fast and easy - we provide optimization reagents with varying concentrations of SDS. Moreover, our Chromatin Shearing Optimization Kits can be used for the optimization of chromatin preparation with our kits for ChIP.</p>
</div>
<div class="small-12 medium-12 large-12 columns">
<div class="page" title="Page 7">
<table>
<tbody>
<tr valign="middle">
<td></td>
<td style="text-align: center;"><strong><a href="https://www.diagenode.com/p/chromatin-shearing-optimization-kit-low-sds-100-million-cells">Chromatin Shearing Kit Low SDS (for Histone)</a></strong></td>
<td style="text-align: center;"><strong><a href="https://www.diagenode.com/p/chromatin-shearing-optimization-kit-low-sds-for-tfs-25-rxns">Chromatin Shearing Kit Low SDS (for TF)</a></strong></td>
<td style="text-align: center;"><strong><a href="https://www.diagenode.com/p/chromatin-shearing-optimization-kit-high-sds-100-million-cells">Chromatin Shearing Kit High SDS</a></strong></td>
<td style="text-align: center;"><strong><a href="https://www.diagenode.com/p/chromatin-shearing-optimization-kit-medium-sds-100-million-cells">Chromatin Shearing Kit (for Plant)</a></strong></td>
</tr>
<tr valign="middle" style="background-color: #fff;">
<td>
<p><strong>SDS concentration</strong></p>
</td>
<td style="text-align: center;">
<p>< 0.1%</p>
</td>
<td style="text-align: center;">
<p>0.2%</p>
</td>
<td style="text-align: center;">
<p>1%</p>
</td>
<td style="text-align: center;">
<p>0.5%</p>
</td>
</tr>
<tr valign="middle" style="background-color: #fff;">
<td>
<p><strong>Nuclei isolation</strong></p>
</td>
<td style="text-align: center;">
<p>Yes</p>
</td>
<td style="text-align: center;">
<p>Yes</p>
</td>
<td style="text-align: center;">
<p>No</p>
</td>
<td style="text-align: center;">
<p>Yes</p>
</td>
</tr>
<tr valign="middle" style="background-color: #fff;">
<td>
<p><strong>Allows for shearing of... cells/tissue</strong></p>
</td>
<td style="text-align: center;">
<p>100 million cells</p>
</td>
<td style="text-align: center;">
<p>100 million cells</p>
</td>
<td style="text-align: center;">
<p>100 million cells</p>
</td>
<td style="text-align: center;">
<p>up to 25 g of tissue</p>
</td>
</tr>
<tr valign="middle" style="background-color: #fff;">
<td>
<p><strong>Corresponding to shearing buffers from</strong></p>
</td>
<td style="text-align: center;">
<p><a href="https://www.diagenode.com/en/p/ideal-chip-seq-kit-x24-24-rxns">iDeal ChIP-seq kit for Histones</a></p>
</td>
<td style="text-align: center;">
<p><a href="https://www.diagenode.com/en/p/ideal-chip-seq-kit-for-transcription-factors-x24-24-rxns">iDeal ChIP-seq Kit for Transcription Factors</a></p>
<p><a href="https://www.diagenode.com/en/p/ideal-chip-qpcr-kit">iDeal ChIP qPCR kit</a></p>
</td>
<td style="text-align: center;">
<p><a href="https://www.diagenode.com/en/p/true-microchip-kit-x16-16-rxns">True MicroChIP kit</a></p>
</td>
<td style="text-align: center;">
<p><a href="https://www.diagenode.com/en/p/universal-plant-chip-seq-kit-x24-24-rxns">Universal Plant ChIP-seq kit</a></p>
</td>
</tr>
</tbody>
</table>
<p><em><span style="font-weight: 400;">Table comes from our </span><a href="https://www.diagenode.com/protocols/bioruptor-pico-chromatin-preparation-guide"><span style="font-weight: 400;">Guide for successful chromatin preparation using the Bioruptor® Pico</span></a></em></p>
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'description' => '<p>Optimal detergent concentration is required for best results with chromatin shearing. Diagenode offers four kits with different SDS concentrations that have been pre-optimized for your specific workflow requirements. Our Chromatin EasyShear Kits (previous name: Chromatin Shearing Optimization Kits) together with the Bioruptor combine efficient cell lysis and chromatin shearing leading to consistent results.</p>
<p>The Chromatin EasyShear Kits are recommended for:</p>
<ul>
<li>The optimization of the chromatin shearing and/or chromatin preparation prior to any ChIP protocol</li>
<li>The optimization of the chromatin shearing of a new cell line/new sample type prior to ChIP using Diagenode’s ChIP kits</li>
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<p>Check all <a href="https://www.diagenode.com/en/categories/chromatin-shearing">Chromatin EasyShear Kits</a>.</p>
<p>Guide for the optimal chromatin preparation using Chromatin EasyShear Kits – <a href="https://www.diagenode.com/en/pages/chromatin-prep-easyshear-kit-guide">Read more</a></p>
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<p><span>The first critical step of a successful Chromatin immunoprecipitation (ChIP) experiment is the preparation of sheared chromatin. We therefore suggest to use one of our optimized shearing ChIP kits.</span></p>
<span>• Chromatin EasyShear Kit - Ultra Low SDS <br />• Chromatin <span>EasyShear Kit</span> <span>-</span> Low SDS <br />• Chromatin EasyShear Kit - High SDS </span></div>
<div class="column"><span>• Chromatin EasyShear Kit for Plant</span><span></span>
<p><span></span></p>
<p><span>Our Chromatin EasyShear kits are used in combination with the Bioruptor</span><span>® </span><span>in order to ensure a highly reproducible chromatin shearing and make sure you obtain the right fragment size needed for your experiment. Establish optimal conditions required for shearing cross-linked chromatin (protein-DNA) is usually laborious; the protocol of the Chromatin EasyShear kits is fast, easy-to-use and optimized to get the best results possible. </span></p>
<p><span>Learn more about Diagenode <a href="https://www.diagenode.com/en/categories/chromatin-shearing">Chromatin EasyShear Kits</a>.</span></p>
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<p>The Diagenode <strong>µChIPmentation Kit for Histones</strong> is optimized to perform ChIP-seq on as little as <strong>10.000 cells</strong> from cell fixation to purified libraries. To <strong>reduce DNA lost</strong> the number of sample transfer from tube to tube has been limited: during whole workflow - from cell fixation to library prurification - only 3 tubes per sample are used. Reduced number of steps, reduced number of sample transfer and <a href="https://www.diagenode.com/en/categories/chromatin-ip-chipmentation" target="_blank">ChIPmentation technology</a> itself enable for <strong>efficient and robust ChIP-seq</strong> on <strong>limited amount of sample</strong>. The kit µChIPmentation for Histones includes all reagents for chromatin preparation, chromatin immunoprecipitation and library preparation using tagmentation. The primer indexes for multiplexing are not included in the kit and have to be purchase separately - <a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set1">read more</a>.</p>
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<li class="accordion-navigation"><a style="background-color: white;" href="#workflow"><i class="fa fa-caret-right"></i> Workflow of µChIPmentation for Histones - Read more</a>
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<p style="text-align: left;"><strong>Figure 1. Comparison between ChIPmentation and µChIPmentation </strong><br />ChIPmentation: chromatin preparation has been performed on 7 M K562 cells using the ChIPmentation Kit for Histones (Cat. No. C01011000) and diluted chromatin from 500.000 cells was used for the immunoprecipitation. µChIPmentation: chromatin preparation and immunoprecipitation have been performed on 10.000 K562 cells using the µChIPmentation Kit for Histones (Cat. No. C01011011) and 24 SI for Tagmented libraries (Cat. No. C010111032). The Diagenode antibody targeting H3K27me3 (Cat. No. C15410195) was used.<br /> A. Distribution of the ChIPmentation and µChIPmentation readsets in a representative region of the genome (in duplicates). B. Comparison of the top 40% peaks from µChIPmentation (10.000 cells) with dataset generated with ChIPmentation (500.000 cells).</p>
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<p><strong>Figure 2. Sequencing profiles of µChIPmentation libraries </strong><br />Chromatin preparation and immunoprecipitation have been performed on 10.000 cells using the µChIPmentation Kit for Histones (Cat. No. C01011011) and 24 SI for Tagmented libraries (Cat. No. C010111032) using K562 cells. The Diagenode antibodies targeting H3K4me3 (Cat. No. C15410003), H3K27ac (Cat. No. C15410196), H3K27me3 (Cat. No. C15410195) and H3K9me3 (Cat. No. C15410193) have been used.</p>
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<p><strong>Figure 3. Integrative genomics viewer (IGV) visualization of from ChIP-seq experiments using H3K27me3 antibody (Diagenode, Cat. No. <span>C15410195</span>) and 10.000 cells of K562 cells per immunoprecipitation.<br /></strong>Cells were FACS-sorted and ChIP has been performed accordingly to µChIPmentation protocol. Batch chromatin preparation followed by immunoprecipitation has been used per comparison as indicated.</p>
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<p><a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies" target="_blank">ChIP-seq grade anti-histone antibodies</a> provide high yields with excellent specificity and sensitivity.</p>
<p>For fast and efficient isolation of magnetic beads we recommend the magnetic racks <a href="https://www.diagenode.com/en/p/diamag02-magnetic-rack-1-unit">DiaMag0.2</a>.</p>
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<p><span>The first critical step of a successful Chromatin immunoprecipitation (ChIP) experiment is the preparation of sheared chromatin. We therefore suggest to use one of our optimized shearing ChIP kits.</span></p>
<span>• Chromatin EasyShear Kit - Ultra Low SDS <br />• Chromatin <span>EasyShear Kit</span> <span>-</span> Low SDS <br />• Chromatin EasyShear Kit - High SDS </span></div>
<div class="column"><span>• Chromatin EasyShear Kit for Plant</span><span></span>
<p><span></span></p>
<p><span>Our Chromatin EasyShear kits are used in combination with the Bioruptor</span><span>® </span><span>in order to ensure a highly reproducible chromatin shearing and make sure you obtain the right fragment size needed for your experiment. Establish optimal conditions required for shearing cross-linked chromatin (protein-DNA) is usually laborious; the protocol of the Chromatin EasyShear kits is fast, easy-to-use and optimized to get the best results possible. </span></p>
<p><span>Learn more about Diagenode <a href="https://www.diagenode.com/en/categories/chromatin-shearing">Chromatin EasyShear Kits</a>.</span></p>
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<p style="text-align: justify;">Previous name: Chromatin Shearing Optimization Kit - High SDS (True Micro ChIP kit)</p>
<p style="text-align: justify;">A high quality chromatin preparation is very complex and requires a lot of optimization. Chromatin EasyShear Kit – High SDS is an optimized solution for efficient chromatin preparation prior to ChIP. The protocol, buffers composition, SDS concentration (1%) is optimized for the preparation of chromatin prior to ChIP on low amount of starting material<b> </b>and it is compatible with Diagenode's <a href="https://www.diagenode.com/en/p/true-microchip-kit-x16-16-rxns">True MicroChIP-seq kit</a> and <a href="https://www.diagenode.com/en/p/uchipmentation-for-histones-24-rxns">µChIPmentation Kit for Histones</a>. The kit has been validated with the Bioruptor ultrasonicator for efficient chromatin shearing, leading to chromatin fragments<span> </span><strong>suitable for ChIP</strong><span> </span>with the preserved<span> </span><strong>epitopes</strong>.</p>
<p style="text-align: justify;">Check all <a href="https://www.diagenode.com/en/categories/chromatin-shearing">Chromatin EasyShear Kits</a>.</p>
<p style="text-align: justify;">Guide for the optimal chromatin preparation using Chromatin EasyShear Kits – <a href="https://www.diagenode.com/en/pages/chromatin-prep-easyshear-kit-guide">Read more</a></p>',
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<p style="text-align: justify;">Previous name: Chromatin Shearing Optimization Kit - High SDS (True Micro ChIP kit)</p>
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<p style="text-align: justify;">Check all <a href="https://www.diagenode.com/en/categories/chromatin-shearing">Chromatin EasyShear Kits</a>.</p>
<p style="text-align: justify;">Guide for the optimal chromatin preparation using Chromatin EasyShear Kits – <a href="https://www.diagenode.com/en/pages/chromatin-prep-easyshear-kit-guide">Read more</a></p>',
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<p>The <b>True </b><b>MicroChIP-seq</b><b> kit </b>provides a robust ChIP protocol suitable for the investigation of histone modifications within chromatin from as few as <b>10 000 cells</b>, including <b>FACS sorted cells</b>. The kit can be used for chromatin preparation for downstream ChIP-qPCR or ChIP-seq analysis. The <b>complete kit</b> contains everything you need for start-to-finish ChIP including all validated buffers and reagents for chromatin shearing, immunoprecipitation and DNA purification for exceptional <strong>ChIP-qPCR</strong> or <strong>ChIP-seq</strong> results. In addition, positive control antibodies and negative control PCR primers are included for your convenience and assurance of result sensitivity and specificity.</p>
<p>The True MicroChIP-seq kit offers unique benefits:</p>
<ul>
<li>An <b>optimized chromatin preparation </b>protocol compatible with low number of cells (<b>10.000</b>) in combination with the Bioruptor™ shearing device</li>
<li>Most <b>complete kit </b>available (covers all steps and includes control antibodies and primers)</li>
<li><b>Magnetic beads </b>make ChIP easy, fast, and more reproducible</li>
<li>MicroChIP DiaPure columns (included in the kit) enable the <b>maximum recovery </b>of immunoprecipitation DNA suitable for any downstream application</li>
<li><b>Excellent </b><b>ChIP</b><b>-seq </b>result when combined with <a href="https://www.diagenode.com/en/categories/library-preparation-for-ChIP-seq">MicroPlex</a><a href="https://www.diagenode.com/en/categories/library-preparation-for-ChIP-seq"> Library Preparation kit </a>adapted for low input</li>
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<p>For fast ChIP-seq on low input – check out Diagenode’s <a href="https://www.diagenode.com/en/p/uchipmentation-for-histones-24-rxns">µ</a><a href="https://www.diagenode.com/en/p/uchipmentation-for-histones-24-rxns">ChIPmentation</a><a href="https://www.diagenode.com/en/p/uchipmentation-for-histones-24-rxns"> for histones</a>.</p>
<p><sub>The True MicroChIP-seq kit, Cat. No. C01010132 is an upgraded version of the kit True MicroChIP, Cat. No. C01010130, with the new validated protocols (e.g. FACS sorted cells) and MicroChIP DiaPure columns included in the kit.</sub></p>',
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<li><b>Revolutionary:</b> Only 10,000 cells needed for complete ChIP-seq procedure</li>
<li><b>Validated on</b> studies for histone marks</li>
<li><b>Automated protocol </b>for the IP-Star<sup>®</sup> Compact Automated Platform available</li>
</ul>
<p></p>
<p>The True MicroChIP-seq kit protocol has been optimized for the use of 10,000 - 100,000 cells per immunoprecipitation reaction. Regarding chromatin immunoprecipitation, three protocol variants have been optimized:<br />starting with a batch, starting with an individual sample and starting with the FACS-sorted cells.</p>
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<div>
<h3>High efficiency ChIP on 10,000 cells</h3>
<div class="large-10 small-12 medium-10 large-centered medium-centered small-centered columns"><img src="https://www.diagenode.com/img/product/kits/true-micro-chip-histone-results.png" width="800px" /></div>
<div class="large-10 small-12 medium-10 large-centered medium-centered small-centered columns"><center>
<p><small><strong>Figure 1. </strong>ChIP efficiency on 10,000 cells. ChIP was performed on human Hela cells using the Diagenode antibodies <a href="https://www.diagenode.com/en/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">H3K4me3</a> (Cat. No. C15410003), <a href="https://www.diagenode.com/en/p/h3k27ac-polyclonal-antibody-classic-50-mg-42-ml">H3K27ac</a> (C15410174), <a href="https://www.diagenode.com/en/p/h3k9me3-polyclonal-antibody-classic-50-ug">H3K9me3</a> (C15410056) and <a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-classic-50-mg-34-ml">H3K27me3</a> (C15410069). Sheared chromatin from 10,000 cells and 0.1 µg (H3K27ac), 0.25 µg (H3K4me3 and H3K27me3) or 0.5 µg (H3K9me3) of the antibody were used per IP. Corresponding amount of IgG was used as control. Quantitative PCR was performed with primers for corresponding positive and negative loci. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
</center></div>
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<div>
<h3>True MicroChIP-seq protocol in a combination with MicroPlex library preparation kit results in reliable and accurate sequencing data</h3>
<div class="large-10 small-12 medium-10 large-centered medium-centered small-centered columns"><img src="https://www.diagenode.com/img/product/kits/fig2-truemicro.jpg" alt="True MicroChip results" width="800px" /></div>
<div class="large-10 small-12 medium-10 large-centered medium-centered small-centered columns"><center>
<p><small><strong>Figure 2.</strong> Integrative genomics viewer (IGV) visualization of ChIP-seq experiments using 50.000 of K562 cells. ChIP has been performed accordingly to True MicroChIP protocol followed by the library preparation using MicroPlex Library Preparation Kit (C05010001). The above figure shows the peaks from ChIP-seq experiments using the following antibodies: H3K4me1 (C15410194), H3K9/14ac (C15410200), H3K27ac (C15410196) and H3K36me3 (C15410192).</small></p>
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<div>
<h3>Successful chromatin profiling from 10.000 of FACS-sorted cells</h3>
<div class="large-10 small-12 medium-10 large-centered medium-centered small-centered columns"><img src="https://www.diagenode.com/img/product/kits/fig3ab-truemicro.jpg" alt="small non coding RNA" width="800px" /></div>
<div class="large-10 small-12 medium-10 large-centered medium-centered small-centered columns"><center>
<p><small><strong>Figure 3.</strong> (A) Integrative genomics viewer (IGV) visualization of ChIP-seq experiments and heatmap 3kb upstream and downstream of the TSS (B) for H3K4me3. ChIP has been performed using 10.000 of FACS-sorted cells (K562) and H3K4me3 antibody (C15410003) accordingly to True MicroChIP protocol followed by the library preparation using MicroPlex Library Preparation Kit (C05010001). Data were compared to ENCODE standards.</small></p>
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'label2' => 'Additional solutions compatible with the True MicroChIP-seq Kit',
'info2' => '<p><span style="font-weight: 400;">The <a href="https://www.diagenode.com/en/p/chromatin-shearing-optimization-kit-high-sds-100-million-cells">Chromatin EasyShear Kit – High SDS</a></span><span style="font-weight: 400;"> Recommended for the optimizing chromatin shearing.</span></p>
<p><a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies"><span style="font-weight: 400;">ChIP-seq grade antibodies</span></a><span style="font-weight: 400;"> for high yields, specificity, and sensitivity.</span></p>
<p><span style="font-weight: 400;">Check the list of available </span><a href="https://www.diagenode.com/en/categories/primer-pairs"><span style="font-weight: 400;">primer pairs</span></a><span style="font-weight: 400;"> designed for high specificity to specific genomic regions.</span></p>
<p><span style="font-weight: 400;">For library preparation of immunoprecipitated samples we recommend to use the </span><b> </b><a href="https://www.diagenode.com/en/categories/library-preparation-for-ChIP-seq"><span style="font-weight: 400;">MicroPlex Library Preparation Kit</span></a><span style="font-weight: 400;"> - validated for library preparation from picogram inputs.</span></p>
<p><span style="font-weight: 400;">For IP-Star Automation users, check out the </span><a href="https://www.diagenode.com/en/p/auto-true-microchip-kit-16-rxns"><span style="font-weight: 400;">automated version</span></a><span style="font-weight: 400;"> of this kit.</span></p>
<p><span style="font-weight: 400;">Application note: </span><a href="https://www.diagenode.com/files/application_notes/Diagenode_AATI_Joint.pdf"><span style="font-weight: 400;">Best Workflow Practices for ChIP-seq Analysis with Small Samples</span></a></p>
<p></p>',
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'info3' => '<p>The True MicroChIP-seq kit is compatible with a broad variety of cell lines, tissues and species - some examples are shown below. Other species / cell lines / tissues can be used with this kit.</p>
<p><strong>Cell lines:</strong></p>
<p>Bovine: blastocysts,<br />Drosophila: embryos, salivary glands<br />Human: EndoC-ẞH1 cells, HeLa cells, PBMC, urothelial cells<br />Mouse: adipocytes, B cells, blastocysts, pre-B cells, BMDM cells, chondrocytes, embryonic stem cells, KH2 cells, LSK cells, macrophages, MEP cells, microglia, NK cells, oocytes, pancreatic cells, P19Cl6 cells, RPE cells,</p>
<p>Other cell lines / species: compatible, not tested</p>
<p><strong>Tissues:</strong></p>
<p>Horse: adipose tissue</p>
<p>Mice: intestine tissue</p>
<p>Other tissues: not tested</p>',
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'description' => '<p>The Auto True MicroChIP kit in combination with the <a href="../p/microplex-library-preparation-kit-v2-x12-12-indices-12-rxns">MicroPlex Library Preparation™ kit</a> allows for performing ChIP-seq on as few as 10,000 cells. This microChIP-seq assay has been validated with the new generation <a href="../p/sx-8g-ip-star-compact-automated-system-1-unit">IP-Star<sup>®</sup> Compact Automated Workstation</a>. Furthermore Diagenode's high quality ChIP-seq grade antibodies have been used.</p>
<div class="row">
<div class="small-12 medium-5 large-6 columns"><small style="color: #666;">Video article</small>
<div class="flex-video"><iframe width="420" height="315" style="float: left; margin-right: 20px;" src="//www.youtube.com/embed/JZC9ARbLh_U?rel=0" frameborder="0" allowfullscreen="allowfullscreen"></iframe></div>
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<div class="small-12 medium-7 large-6 columns">
<h3>Automating ChIP-seq Experiments to Generate Epigenetic Profiles on 10,000 HeLa Cells</h3>
<p><a href="https://www.youtube.com/watch?v=JZC9ARbLh_U"><img src="https://www.diagenode.com/img/jove_logo.png" alt="Jove logo" border="0" width="100" height="58" /></a></p>
</div>
</div>',
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'label2' => '特徴',
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<li><strong>Revolutionary:</strong> Only 10.000 cells needed for complete ChIP-seq procedure</li>
<li><strong>Reliable and accurate</strong> sequencing library preparation from just picogram inputs</li>
<li><strong>Validated on</strong> studies for histone marks and with the IP-Star<sup>®</sup> Compact Automated Platform</li>
</ul>
<p><img src="https://www.diagenode.com/img/product/kits/true-micro-chip-histone-results.png" alt="ChIP on 10,000 cells" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><strong>High efficiency ChIP on 10,000 cells</strong><br />ChIP efficiency on 10 000 cells. ChIP was performed on human Hela cells using the Diagenode antibodies H3K4me3 (Cat. No. pAb-003-050), H3K27ac (pAb-174-050), H3K9me3 (pAb-056-050) and H3K27me3 (pAb-069-050). Sheared chromatin from 10 000 cells and 0.1 µg (H3K27ac), 0.25 µg (H3K4me3 and H3K27me3) or 0.5 µg (H3K9me3) of the antibody were used per IP. Corresponding amount of IgG was used as control. Quantitative PCR was performed with primers for corresponding positive and negative loci. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</p>
<p><img src="https://www.diagenode.com/img/product/kits/true-micro-chip-reliable-A.png" alt="ChIP amplification" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/kits/true-micro-chip-reliable-B.png" alt="Match between True MicroChIP peaks and the reference dataset" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><strong>Reliable detection of enrichments in ChIP-seq</strong><br />A: ChIP has been peformed with H3K4me3 antibody, amplification of 17 pg of DNA ChIP'd from 10.000 cells and amplification of 35 pg of DNA ChIP'd from 100.000 cells (control experiment). The IP'd DNA was amplified and transformed into a sequencing-ready preparation for the Illumina plateform with the MicroPlex Library Preparation kit. The library was then analysed on an Illumina<sup>®</sup> Genome Analyzer. Cluster generation and sequencing were performed according to the manufacturer's instructions.<br />B: We observed a perfect match between the top 40% of True MicroChIP peaks and the reference dataset. Based on the NIH Encode project criterion, ChIP-seq results are considered reproducible between an original and reproduced dataset if the top 40% of peaks have at least an 80% overlap ratio with the compared dataset.</p>
<p><img src="https://www.diagenode.com/img/product/kits/true-micro-chip-result.png" alt="Cell lysis" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><strong>Hela cells were fixed with 1% formaldehyde (for 10 minutes at RT)</strong><br /> Cell lysis was performed using the Lysis Buffer tL1 of the Diagenode True MicroChIP kit. Samples corresponding to 10 000 cells are sheared during 5 rounds of 5 cycles of 30 seconds “ON” / 30 seconds “OFF” with the Bioruptor<sup>®</sup> Plus combined with the Bioruptor<sup>®</sup> Water cooler (Cat No. BioAcc-cool) at HIGH power setting (position H). For optimal results, samples are vortexed before and after performing 5 sonication cycles, followed by a short centrifugation at 4°C. 10 μl of DNA (equivalent to 60 000 cells) are analysed on a 1.5% agarose gel.</p>',
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'slug' => 'auto-true-microchip-kit-16-rxns',
'meta_title' => 'Auto True MicroChIP kit',
'meta_keywords' => '',
'meta_description' => 'Auto True MicroChIP kit',
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'name' => 'µChIPmentation Kit for Histones',
'description' => '<p><a href="https://www.diagenode.com/files/products/kits/microchipmentation-for-histones.pdf"><img src="https://www.diagenode.com/img/buttons/bt-manual.png" /></a></p>
<p>The Diagenode <strong>µChIPmentation Kit for Histones</strong> is optimized to perform ChIP-seq on as little as <strong>10.000 cells</strong> from cell fixation to purified libraries. To <strong>reduce DNA lost</strong> the number of sample transfer from tube to tube has been limited: during whole workflow - from cell fixation to library prurification - only 3 tubes per sample are used. Reduced number of steps, reduced number of sample transfer and <a href="https://www.diagenode.com/en/categories/chromatin-ip-chipmentation" target="_blank">ChIPmentation technology</a> itself enable for <strong>efficient and robust ChIP-seq</strong> on <strong>limited amount of sample</strong>. The kit µChIPmentation for Histones includes all reagents for chromatin preparation, chromatin immunoprecipitation and library preparation using tagmentation. The primer indexes for multiplexing are not included in the kit and have to be purchase separately - <a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set1">read more</a>.</p>
<div class="extra-spaced">
<h3>Benefits of the µChIPmentation system for histone ChIP-seq</h3>
<ul>
<li><strong>Easier</strong> and <strong>faster</strong> than classical ChIP-seq</li>
<li>Optimized for <strong>low input samples</strong>: as little as 10,000 cells</li>
<li>Protocol optimized for <strong>FACS-sorted cells</strong></li>
<li>Validated for various <strong>histone marks</strong></li>
<li><strong>High quality</strong> sequencing data</li>
</ul>
</div>',
'label1' => 'Characteristics',
'info1' => '<p>ChIPmentation is based on tagmentation that allows library preparation to be integrated during the ChIP itself using transposase and sequencing-compatible adaptors. Our new improved µChIPmentation protocol combines 3 features for guaranteeing high quality sequencing data on small sample inputs 1) optimized chromatin shearing preparation protocol, 2) reduced number of steps, 3) reduced number of sample transfer from tube to tube - only 3 tubes per sample for the whole process, from cell fixation to purified libraries.</p>
<ul class="accordion" data-accordion="" style="margin-left: 0;">
<li class="accordion-navigation"><a style="background-color: white;" href="#workflow"><i class="fa fa-caret-right"></i> Workflow of µChIPmentation for Histones - Read more</a>
<div id="workflow" class="content">
<div class="extra-spaced"><center><img src="https://www.diagenode.com/img/product/kits/workflow-micro-chipmentation.png" /></center></div>
</div>
</li>
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<div class="row extra-spaced">
<div class="small-12 medium-12 large-12 columns"><strong>A.</strong><img src="https://www.diagenode.com/img/product/kits/micro-chipmentation-fig1A.png" /></div>
</div>
<div class="row extra-spaced">
<div class="small-12 medium-5 large-5 columns" columns=""><strong>B.</strong><img src="https://www.diagenode.com/img/product/kits/micro-chipmentation-fig1B.png" /></div>
<div class="small-12 medium-7 large-7 columns">
<p style="text-align: left;"><strong>Figure 1. Comparison between ChIPmentation and µChIPmentation </strong><br />ChIPmentation: chromatin preparation has been performed on 7 M K562 cells using the ChIPmentation Kit for Histones (Cat. No. C01011000) and diluted chromatin from 500.000 cells was used for the immunoprecipitation. µChIPmentation: chromatin preparation and immunoprecipitation have been performed on 10.000 K562 cells using the µChIPmentation Kit for Histones (Cat. No. C01011011) and 24 SI for Tagmented libraries (Cat. No. C010111032). The Diagenode antibody targeting H3K27me3 (Cat. No. C15410195) was used.<br /> A. Distribution of the ChIPmentation and µChIPmentation readsets in a representative region of the genome (in duplicates). B. Comparison of the top 40% peaks from µChIPmentation (10.000 cells) with dataset generated with ChIPmentation (500.000 cells).</p>
</div>
</div>
<div class="extra-spaced">
<div class="row">
<div class="small-12 medium-12 large-12 columns"><img src="https://www.diagenode.com/img/product/kits/micro-chipmentation-fig2.png" /></div>
</div>
<div class="row">
<div class="small-12 medium-12 large-12 columns">
<p><strong>Figure 2. Sequencing profiles of µChIPmentation libraries </strong><br />Chromatin preparation and immunoprecipitation have been performed on 10.000 cells using the µChIPmentation Kit for Histones (Cat. No. C01011011) and 24 SI for Tagmented libraries (Cat. No. C010111032) using K562 cells. The Diagenode antibodies targeting H3K4me3 (Cat. No. C15410003), H3K27ac (Cat. No. C15410196), H3K27me3 (Cat. No. C15410195) and H3K9me3 (Cat. No. C15410193) have been used.</p>
</div>
</div>
</div>
<div class="extra-spaced">
<div class="row">
<div class="small-12 medium-12 large-12 columns"><img src="https://www.diagenode.com/img/product/kits/micro-chipmentation-fig3.png" /></div>
</div>
<div class="row">
<div class="small-12 medium-12 large-12 columns">
<p><strong>Figure 3. Integrative genomics viewer (IGV) visualization of from ChIP-seq experiments using H3K27me3 antibody (Diagenode, Cat. No. <span>C15410195</span>) and 10.000 cells of K562 cells per immunoprecipitation.<br /></strong>Cells were FACS-sorted and ChIP has been performed accordingly to µChIPmentation protocol. Batch chromatin preparation followed by immunoprecipitation has been used per comparison as indicated.</p>
</div>
</div>
</div>',
'label2' => 'Additional solutions for µChIPmentation Kit for Histones',
'info2' => '<p><a href="https://www.diagenode.com/en/p/chromatin-shearing-optimization-kit-high-sds-100-million-cells">Chromatin EasyShear Kit - High SDS</a> optimizes chromatin shearing, a critical step for ChIP.</p>
<p><a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies" target="_blank">ChIP-seq grade anti-histone antibodies</a> provide high yields with excellent specificity and sensitivity.</p>
<p>For fast and efficient isolation of magnetic beads we recommend the magnetic racks <a href="https://www.diagenode.com/en/p/diamag02-magnetic-rack-1-unit">DiaMag0.2</a>.</p>
<p>Primer indexes for multiplexing:</p>
<ul class="no-bullet">
<li><a href="https://www.diagenode.com/en/p/24-si-for-tagmented-libraries" target="_blank">24 SI for Tagmented libraries Cat. No. C01011032</a></li>
<li><a href="https://www.diagenode.com/en/p/8-si-for-tagmented-libraries" target="_blank">8 SI for Tagmented libraries Cat. No. C01011033</a></li>
<li><em></em><a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set1">24 UDI for Tagmented libraries - Set I, Cat. No. C0101134</a></li>
<li><a href="https://www.diagenode.com/en/p/8-unique-dual-indexes-for-tagmented-libraries">8 UDI for Tagmented libraries, Cat. No. C0101135</a></li>
<li><a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set2">24 UDI for Tagmented libraries - Set II, Cat. No. C0101136</a></li>
<li><a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set3">24 UDI for Tagmented libraries - Set III, Cat. No. C0101137</a></li>
<li></li>
</ul>',
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'description' => '<div class="row">
<div class="small-12 medium-12 large-12 columns">The most important steps for a successful ChIP include both cell fixation and lysis, and chromatin shearing. Researchers often overlook the critical nature of both of these steps. Eliminating inconsistencies in the shearing step, <strong>Diagenode's Bioruptor</strong><sup>®</sup> uses state-of-the-art ultrasound <strong>ACT</strong> (<strong>A</strong>daptive <strong>C</strong>avitation <strong>T</strong>echnology) to efficiently shear chromatin. ACT enables the highest chromatin quality for high IP efficiency and sensitivity for ChIP experiments with gentle yet highly effective shearing forces. Additionally, the Bioruptor<sup>®</sup> provides a precisely controlled temperature environment that preserves chromatin from heat degradation such that protein-DNA complexes are well-preserved for sensitive, unbiased, and accurate ChIP.<br /><br /> <strong>Diagenode's Bioruptor</strong><sup>®</sup> is the instrument of choice for chromatin shearing used for a number of downstream applications such as qPCR and ChIP-seq that require optimally sheared, unbiased chromatin.</div>
<div class="small-12 medium-12 large-12 columns text-center"><br /><img src="https://www.diagenode.com/img/applications/pico_dna_shearing_fig2.png" /></div>
<div class="small-10 medium-10 large-10 columns end small-offset-1"><small> <br /><strong>Panel A, 10 µl volume:</strong> Chromatin samples are sheared for 10, 20 and 30 cycles of 30 sec ON/30 sec OFF with the Bioruptor® Pico using 0.1 ml Bioruptor® Microtubes (Cat. No. B01200041). <strong>Panel B, 100 µl volume:</strong> Chromatin samples are sheared for 10 cycles of 30 sec ON/30 sec OFF with the Bioruptor® Pico using 0.65 ml Bioruptor® Microtubes (Cat. No. WA-005-0500). <strong>Panel C, 300 µl volume:</strong> Chromatin samples are sheared for 5, 10 and 15 cycles of 30 sec ON/30 sec OFF with the Bioruptor® Pico using using 1.5 ml Bioruptor microtubes (Cat. No. C30010016). Prior to de-crosslinking, samples are treated with RNase cocktail mixture at 37°C during 1 hour. The sheared chromatin is then de-crosslinked overnight and phenol/chloroform purified as described in the kit manual. 10 µl of DNA (equivalent of 500, 000 cells) are analyzed on a 2% agarose gel (MW corresponds to the 100 bp DNA molecular weight marker).</small></div>
<div class="small-12 medium-12 large-12 columns"><br /><br /></div>
<div class="small-12 medium-12 large-12 columns">
<p>It is important to establish optimal conditions to shear crosslinked chromatin to get the correct fragment sizes needed for ChIP. Usually this process requires both optimizing sonication conditions as well as optimizing SDS concentration, which is laborious. With the Chromatin Shearing Optimization Kits, optimization is fast and easy - we provide optimization reagents with varying concentrations of SDS. Moreover, our Chromatin Shearing Optimization Kits can be used for the optimization of chromatin preparation with our kits for ChIP.</p>
</div>
<div class="small-12 medium-12 large-12 columns">
<div class="page" title="Page 7">
<table>
<tbody>
<tr valign="middle">
<td></td>
<td style="text-align: center;"><strong><a href="https://www.diagenode.com/p/chromatin-shearing-optimization-kit-low-sds-100-million-cells">Chromatin Shearing Kit Low SDS (for Histone)</a></strong></td>
<td style="text-align: center;"><strong><a href="https://www.diagenode.com/p/chromatin-shearing-optimization-kit-low-sds-for-tfs-25-rxns">Chromatin Shearing Kit Low SDS (for TF)</a></strong></td>
<td style="text-align: center;"><strong><a href="https://www.diagenode.com/p/chromatin-shearing-optimization-kit-high-sds-100-million-cells">Chromatin Shearing Kit High SDS</a></strong></td>
<td style="text-align: center;"><strong><a href="https://www.diagenode.com/p/chromatin-shearing-optimization-kit-medium-sds-100-million-cells">Chromatin Shearing Kit (for Plant)</a></strong></td>
</tr>
<tr valign="middle" style="background-color: #fff;">
<td>
<p><strong>SDS concentration</strong></p>
</td>
<td style="text-align: center;">
<p>< 0.1%</p>
</td>
<td style="text-align: center;">
<p>0.2%</p>
</td>
<td style="text-align: center;">
<p>1%</p>
</td>
<td style="text-align: center;">
<p>0.5%</p>
</td>
</tr>
<tr valign="middle" style="background-color: #fff;">
<td>
<p><strong>Nuclei isolation</strong></p>
</td>
<td style="text-align: center;">
<p>Yes</p>
</td>
<td style="text-align: center;">
<p>Yes</p>
</td>
<td style="text-align: center;">
<p>No</p>
</td>
<td style="text-align: center;">
<p>Yes</p>
</td>
</tr>
<tr valign="middle" style="background-color: #fff;">
<td>
<p><strong>Allows for shearing of... cells/tissue</strong></p>
</td>
<td style="text-align: center;">
<p>100 million cells</p>
</td>
<td style="text-align: center;">
<p>100 million cells</p>
</td>
<td style="text-align: center;">
<p>100 million cells</p>
</td>
<td style="text-align: center;">
<p>up to 25 g of tissue</p>
</td>
</tr>
<tr valign="middle" style="background-color: #fff;">
<td>
<p><strong>Corresponding to shearing buffers from</strong></p>
</td>
<td style="text-align: center;">
<p><a href="https://www.diagenode.com/en/p/ideal-chip-seq-kit-x24-24-rxns">iDeal ChIP-seq kit for Histones</a></p>
</td>
<td style="text-align: center;">
<p><a href="https://www.diagenode.com/en/p/ideal-chip-seq-kit-for-transcription-factors-x24-24-rxns">iDeal ChIP-seq Kit for Transcription Factors</a></p>
<p><a href="https://www.diagenode.com/en/p/ideal-chip-qpcr-kit">iDeal ChIP qPCR kit</a></p>
</td>
<td style="text-align: center;">
<p><a href="https://www.diagenode.com/en/p/true-microchip-kit-x16-16-rxns">True MicroChIP kit</a></p>
</td>
<td style="text-align: center;">
<p><a href="https://www.diagenode.com/en/p/universal-plant-chip-seq-kit-x24-24-rxns">Universal Plant ChIP-seq kit</a></p>
</td>
</tr>
</tbody>
</table>
<p><em><span style="font-weight: 400;">Table comes from our </span><a href="https://www.diagenode.com/protocols/bioruptor-pico-chromatin-preparation-guide"><span style="font-weight: 400;">Guide for successful chromatin preparation using the Bioruptor® Pico</span></a></em></p>
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'meta_description' => 'Diagenode's Bioruptor® is the instrument of choice for chromatin shearing used for a number of downstream applications such as qPCR and ChIP-seq that require optimally sheared, unbiased chromatin.',
'meta_title' => 'Chromatin shearing using Bioruptor® sonication device | Diagenode',
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'description' => '<p>Optimal detergent concentration is required for best results with chromatin shearing. Diagenode offers four kits with different SDS concentrations that have been pre-optimized for your specific workflow requirements. Our Chromatin EasyShear Kits (previous name: Chromatin Shearing Optimization Kits) together with the Bioruptor combine efficient cell lysis and chromatin shearing leading to consistent results.</p>
<p>The Chromatin EasyShear Kits are recommended for:</p>
<ul>
<li>The optimization of the chromatin shearing and/or chromatin preparation prior to any ChIP protocol</li>
<li>The optimization of the chromatin shearing of a new cell line/new sample type prior to ChIP using Diagenode’s ChIP kits</li>
</ul>
<p>Check all <a href="https://www.diagenode.com/en/categories/chromatin-shearing">Chromatin EasyShear Kits</a>.</p>
<p>Guide for the optimal chromatin preparation using Chromatin EasyShear Kits – <a href="https://www.diagenode.com/en/pages/chromatin-prep-easyshear-kit-guide">Read more</a></p>
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<p><span>The first critical step of a successful Chromatin immunoprecipitation (ChIP) experiment is the preparation of sheared chromatin. We therefore suggest to use one of our optimized shearing ChIP kits.</span></p>
<span>• Chromatin EasyShear Kit - Ultra Low SDS <br />• Chromatin <span>EasyShear Kit</span> <span>-</span> Low SDS <br />• Chromatin EasyShear Kit - High SDS </span></div>
<div class="column"><span>• Chromatin EasyShear Kit for Plant</span><span></span>
<p><span></span></p>
<p><span>Our Chromatin EasyShear kits are used in combination with the Bioruptor</span><span>® </span><span>in order to ensure a highly reproducible chromatin shearing and make sure you obtain the right fragment size needed for your experiment. Establish optimal conditions required for shearing cross-linked chromatin (protein-DNA) is usually laborious; the protocol of the Chromatin EasyShear kits is fast, easy-to-use and optimized to get the best results possible. </span></p>
<p><span>Learn more about Diagenode <a href="https://www.diagenode.com/en/categories/chromatin-shearing">Chromatin EasyShear Kits</a>.</span></p>
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<p>The Diagenode <strong>µChIPmentation Kit for Histones</strong> is optimized to perform ChIP-seq on as little as <strong>10.000 cells</strong> from cell fixation to purified libraries. To <strong>reduce DNA lost</strong> the number of sample transfer from tube to tube has been limited: during whole workflow - from cell fixation to library prurification - only 3 tubes per sample are used. Reduced number of steps, reduced number of sample transfer and <a href="https://www.diagenode.com/en/categories/chromatin-ip-chipmentation" target="_blank">ChIPmentation technology</a> itself enable for <strong>efficient and robust ChIP-seq</strong> on <strong>limited amount of sample</strong>. The kit µChIPmentation for Histones includes all reagents for chromatin preparation, chromatin immunoprecipitation and library preparation using tagmentation. The primer indexes for multiplexing are not included in the kit and have to be purchase separately - <a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set1">read more</a>.</p>
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<ul class="accordion" data-accordion="" style="margin-left: 0;">
<li class="accordion-navigation"><a style="background-color: white;" href="#workflow"><i class="fa fa-caret-right"></i> Workflow of µChIPmentation for Histones - Read more</a>
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<p style="text-align: left;"><strong>Figure 1. Comparison between ChIPmentation and µChIPmentation </strong><br />ChIPmentation: chromatin preparation has been performed on 7 M K562 cells using the ChIPmentation Kit for Histones (Cat. No. C01011000) and diluted chromatin from 500.000 cells was used for the immunoprecipitation. µChIPmentation: chromatin preparation and immunoprecipitation have been performed on 10.000 K562 cells using the µChIPmentation Kit for Histones (Cat. No. C01011011) and 24 SI for Tagmented libraries (Cat. No. C010111032). The Diagenode antibody targeting H3K27me3 (Cat. No. C15410195) was used.<br /> A. Distribution of the ChIPmentation and µChIPmentation readsets in a representative region of the genome (in duplicates). B. Comparison of the top 40% peaks from µChIPmentation (10.000 cells) with dataset generated with ChIPmentation (500.000 cells).</p>
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<p><strong>Figure 2. Sequencing profiles of µChIPmentation libraries </strong><br />Chromatin preparation and immunoprecipitation have been performed on 10.000 cells using the µChIPmentation Kit for Histones (Cat. No. C01011011) and 24 SI for Tagmented libraries (Cat. No. C010111032) using K562 cells. The Diagenode antibodies targeting H3K4me3 (Cat. No. C15410003), H3K27ac (Cat. No. C15410196), H3K27me3 (Cat. No. C15410195) and H3K9me3 (Cat. No. C15410193) have been used.</p>
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<p><strong>Figure 3. Integrative genomics viewer (IGV) visualization of from ChIP-seq experiments using H3K27me3 antibody (Diagenode, Cat. No. <span>C15410195</span>) and 10.000 cells of K562 cells per immunoprecipitation.<br /></strong>Cells were FACS-sorted and ChIP has been performed accordingly to µChIPmentation protocol. Batch chromatin preparation followed by immunoprecipitation has been used per comparison as indicated.</p>
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<p><a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies" target="_blank">ChIP-seq grade anti-histone antibodies</a> provide high yields with excellent specificity and sensitivity.</p>
<p>For fast and efficient isolation of magnetic beads we recommend the magnetic racks <a href="https://www.diagenode.com/en/p/diamag02-magnetic-rack-1-unit">DiaMag0.2</a>.</p>
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<li><a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set3">24 UDI for Tagmented libraries - Set III, Cat. No. C0101137</a></li>
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<p><span>The first critical step of a successful Chromatin immunoprecipitation (ChIP) experiment is the preparation of sheared chromatin. We therefore suggest to use one of our optimized shearing ChIP kits.</span></p>
<span>• Chromatin EasyShear Kit - Ultra Low SDS <br />• Chromatin <span>EasyShear Kit</span> <span>-</span> Low SDS <br />• Chromatin EasyShear Kit - High SDS </span></div>
<div class="column"><span>• Chromatin EasyShear Kit for Plant</span><span></span>
<p><span></span></p>
<p><span>Our Chromatin EasyShear kits are used in combination with the Bioruptor</span><span>® </span><span>in order to ensure a highly reproducible chromatin shearing and make sure you obtain the right fragment size needed for your experiment. Establish optimal conditions required for shearing cross-linked chromatin (protein-DNA) is usually laborious; the protocol of the Chromatin EasyShear kits is fast, easy-to-use and optimized to get the best results possible. </span></p>
<p><span>Learn more about Diagenode <a href="https://www.diagenode.com/en/categories/chromatin-shearing">Chromatin EasyShear Kits</a>.</span></p>
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<p style="text-align: justify;">Previous name: Chromatin Shearing Optimization Kit - High SDS (True Micro ChIP kit)</p>
<p style="text-align: justify;">A high quality chromatin preparation is very complex and requires a lot of optimization. Chromatin EasyShear Kit – High SDS is an optimized solution for efficient chromatin preparation prior to ChIP. The protocol, buffers composition, SDS concentration (1%) is optimized for the preparation of chromatin prior to ChIP on low amount of starting material<b> </b>and it is compatible with Diagenode's <a href="https://www.diagenode.com/en/p/true-microchip-kit-x16-16-rxns">True MicroChIP-seq kit</a> and <a href="https://www.diagenode.com/en/p/uchipmentation-for-histones-24-rxns">µChIPmentation Kit for Histones</a>. The kit has been validated with the Bioruptor ultrasonicator for efficient chromatin shearing, leading to chromatin fragments<span> </span><strong>suitable for ChIP</strong><span> </span>with the preserved<span> </span><strong>epitopes</strong>.</p>
<p style="text-align: justify;">Check all <a href="https://www.diagenode.com/en/categories/chromatin-shearing">Chromatin EasyShear Kits</a>.</p>
<p style="text-align: justify;">Guide for the optimal chromatin preparation using Chromatin EasyShear Kits – <a href="https://www.diagenode.com/en/pages/chromatin-prep-easyshear-kit-guide">Read more</a></p>',
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<p style="text-align: justify;">Previous name: Chromatin Shearing Optimization Kit - High SDS (True Micro ChIP kit)</p>
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<p style="text-align: justify;">Check all <a href="https://www.diagenode.com/en/categories/chromatin-shearing">Chromatin EasyShear Kits</a>.</p>
<p style="text-align: justify;">Guide for the optimal chromatin preparation using Chromatin EasyShear Kits – <a href="https://www.diagenode.com/en/pages/chromatin-prep-easyshear-kit-guide">Read more</a></p>',
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<p>The <b>True </b><b>MicroChIP-seq</b><b> kit </b>provides a robust ChIP protocol suitable for the investigation of histone modifications within chromatin from as few as <b>10 000 cells</b>, including <b>FACS sorted cells</b>. The kit can be used for chromatin preparation for downstream ChIP-qPCR or ChIP-seq analysis. The <b>complete kit</b> contains everything you need for start-to-finish ChIP including all validated buffers and reagents for chromatin shearing, immunoprecipitation and DNA purification for exceptional <strong>ChIP-qPCR</strong> or <strong>ChIP-seq</strong> results. In addition, positive control antibodies and negative control PCR primers are included for your convenience and assurance of result sensitivity and specificity.</p>
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<li>Most <b>complete kit </b>available (covers all steps and includes control antibodies and primers)</li>
<li><b>Magnetic beads </b>make ChIP easy, fast, and more reproducible</li>
<li>MicroChIP DiaPure columns (included in the kit) enable the <b>maximum recovery </b>of immunoprecipitation DNA suitable for any downstream application</li>
<li><b>Excellent </b><b>ChIP</b><b>-seq </b>result when combined with <a href="https://www.diagenode.com/en/categories/library-preparation-for-ChIP-seq">MicroPlex</a><a href="https://www.diagenode.com/en/categories/library-preparation-for-ChIP-seq"> Library Preparation kit </a>adapted for low input</li>
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<p>For fast ChIP-seq on low input – check out Diagenode’s <a href="https://www.diagenode.com/en/p/uchipmentation-for-histones-24-rxns">µ</a><a href="https://www.diagenode.com/en/p/uchipmentation-for-histones-24-rxns">ChIPmentation</a><a href="https://www.diagenode.com/en/p/uchipmentation-for-histones-24-rxns"> for histones</a>.</p>
<p><sub>The True MicroChIP-seq kit, Cat. No. C01010132 is an upgraded version of the kit True MicroChIP, Cat. No. C01010130, with the new validated protocols (e.g. FACS sorted cells) and MicroChIP DiaPure columns included in the kit.</sub></p>',
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<h3>High efficiency ChIP on 10,000 cells</h3>
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<p><small><strong>Figure 1. </strong>ChIP efficiency on 10,000 cells. ChIP was performed on human Hela cells using the Diagenode antibodies <a href="https://www.diagenode.com/en/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">H3K4me3</a> (Cat. No. C15410003), <a href="https://www.diagenode.com/en/p/h3k27ac-polyclonal-antibody-classic-50-mg-42-ml">H3K27ac</a> (C15410174), <a href="https://www.diagenode.com/en/p/h3k9me3-polyclonal-antibody-classic-50-ug">H3K9me3</a> (C15410056) and <a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-classic-50-mg-34-ml">H3K27me3</a> (C15410069). Sheared chromatin from 10,000 cells and 0.1 µg (H3K27ac), 0.25 µg (H3K4me3 and H3K27me3) or 0.5 µg (H3K9me3) of the antibody were used per IP. Corresponding amount of IgG was used as control. Quantitative PCR was performed with primers for corresponding positive and negative loci. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<h3>True MicroChIP-seq protocol in a combination with MicroPlex library preparation kit results in reliable and accurate sequencing data</h3>
<div class="large-10 small-12 medium-10 large-centered medium-centered small-centered columns"><img src="https://www.diagenode.com/img/product/kits/fig2-truemicro.jpg" alt="True MicroChip results" width="800px" /></div>
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<p><small><strong>Figure 2.</strong> Integrative genomics viewer (IGV) visualization of ChIP-seq experiments using 50.000 of K562 cells. ChIP has been performed accordingly to True MicroChIP protocol followed by the library preparation using MicroPlex Library Preparation Kit (C05010001). The above figure shows the peaks from ChIP-seq experiments using the following antibodies: H3K4me1 (C15410194), H3K9/14ac (C15410200), H3K27ac (C15410196) and H3K36me3 (C15410192).</small></p>
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<h3>Successful chromatin profiling from 10.000 of FACS-sorted cells</h3>
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<div class="large-10 small-12 medium-10 large-centered medium-centered small-centered columns"><center>
<p><small><strong>Figure 3.</strong> (A) Integrative genomics viewer (IGV) visualization of ChIP-seq experiments and heatmap 3kb upstream and downstream of the TSS (B) for H3K4me3. ChIP has been performed using 10.000 of FACS-sorted cells (K562) and H3K4me3 antibody (C15410003) accordingly to True MicroChIP protocol followed by the library preparation using MicroPlex Library Preparation Kit (C05010001). Data were compared to ENCODE standards.</small></p>
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'info2' => '<p><span style="font-weight: 400;">The <a href="https://www.diagenode.com/en/p/chromatin-shearing-optimization-kit-high-sds-100-million-cells">Chromatin EasyShear Kit – High SDS</a></span><span style="font-weight: 400;"> Recommended for the optimizing chromatin shearing.</span></p>
<p><a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies"><span style="font-weight: 400;">ChIP-seq grade antibodies</span></a><span style="font-weight: 400;"> for high yields, specificity, and sensitivity.</span></p>
<p><span style="font-weight: 400;">Check the list of available </span><a href="https://www.diagenode.com/en/categories/primer-pairs"><span style="font-weight: 400;">primer pairs</span></a><span style="font-weight: 400;"> designed for high specificity to specific genomic regions.</span></p>
<p><span style="font-weight: 400;">For library preparation of immunoprecipitated samples we recommend to use the </span><b> </b><a href="https://www.diagenode.com/en/categories/library-preparation-for-ChIP-seq"><span style="font-weight: 400;">MicroPlex Library Preparation Kit</span></a><span style="font-weight: 400;"> - validated for library preparation from picogram inputs.</span></p>
<p><span style="font-weight: 400;">For IP-Star Automation users, check out the </span><a href="https://www.diagenode.com/en/p/auto-true-microchip-kit-16-rxns"><span style="font-weight: 400;">automated version</span></a><span style="font-weight: 400;"> of this kit.</span></p>
<p><span style="font-weight: 400;">Application note: </span><a href="https://www.diagenode.com/files/application_notes/Diagenode_AATI_Joint.pdf"><span style="font-weight: 400;">Best Workflow Practices for ChIP-seq Analysis with Small Samples</span></a></p>
<p></p>',
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<p><strong>Cell lines:</strong></p>
<p>Bovine: blastocysts,<br />Drosophila: embryos, salivary glands<br />Human: EndoC-ẞH1 cells, HeLa cells, PBMC, urothelial cells<br />Mouse: adipocytes, B cells, blastocysts, pre-B cells, BMDM cells, chondrocytes, embryonic stem cells, KH2 cells, LSK cells, macrophages, MEP cells, microglia, NK cells, oocytes, pancreatic cells, P19Cl6 cells, RPE cells,</p>
<p>Other cell lines / species: compatible, not tested</p>
<p><strong>Tissues:</strong></p>
<p>Horse: adipose tissue</p>
<p>Mice: intestine tissue</p>
<p>Other tissues: not tested</p>',
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'name' => 'Auto True MicroChIP kit',
'description' => '<p>The Auto True MicroChIP kit in combination with the <a href="../p/microplex-library-preparation-kit-v2-x12-12-indices-12-rxns">MicroPlex Library Preparation™ kit</a> allows for performing ChIP-seq on as few as 10,000 cells. This microChIP-seq assay has been validated with the new generation <a href="../p/sx-8g-ip-star-compact-automated-system-1-unit">IP-Star<sup>®</sup> Compact Automated Workstation</a>. Furthermore Diagenode's high quality ChIP-seq grade antibodies have been used.</p>
<div class="row">
<div class="small-12 medium-5 large-6 columns"><small style="color: #666;">Video article</small>
<div class="flex-video"><iframe width="420" height="315" style="float: left; margin-right: 20px;" src="//www.youtube.com/embed/JZC9ARbLh_U?rel=0" frameborder="0" allowfullscreen="allowfullscreen"></iframe></div>
</div>
<div class="small-12 medium-7 large-6 columns">
<h3>Automating ChIP-seq Experiments to Generate Epigenetic Profiles on 10,000 HeLa Cells</h3>
<p><a href="https://www.youtube.com/watch?v=JZC9ARbLh_U"><img src="https://www.diagenode.com/img/jove_logo.png" alt="Jove logo" border="0" width="100" height="58" /></a></p>
</div>
</div>',
'label1' => '',
'info1' => '',
'label2' => '特徴',
'info2' => '<ul>
<li><strong>Revolutionary:</strong> Only 10.000 cells needed for complete ChIP-seq procedure</li>
<li><strong>Reliable and accurate</strong> sequencing library preparation from just picogram inputs</li>
<li><strong>Validated on</strong> studies for histone marks and with the IP-Star<sup>®</sup> Compact Automated Platform</li>
</ul>
<p><img src="https://www.diagenode.com/img/product/kits/true-micro-chip-histone-results.png" alt="ChIP on 10,000 cells" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><strong>High efficiency ChIP on 10,000 cells</strong><br />ChIP efficiency on 10 000 cells. ChIP was performed on human Hela cells using the Diagenode antibodies H3K4me3 (Cat. No. pAb-003-050), H3K27ac (pAb-174-050), H3K9me3 (pAb-056-050) and H3K27me3 (pAb-069-050). Sheared chromatin from 10 000 cells and 0.1 µg (H3K27ac), 0.25 µg (H3K4me3 and H3K27me3) or 0.5 µg (H3K9me3) of the antibody were used per IP. Corresponding amount of IgG was used as control. Quantitative PCR was performed with primers for corresponding positive and negative loci. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</p>
<p><img src="https://www.diagenode.com/img/product/kits/true-micro-chip-reliable-A.png" alt="ChIP amplification" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><img src="https://www.diagenode.com/img/product/kits/true-micro-chip-reliable-B.png" alt="Match between True MicroChIP peaks and the reference dataset" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><strong>Reliable detection of enrichments in ChIP-seq</strong><br />A: ChIP has been peformed with H3K4me3 antibody, amplification of 17 pg of DNA ChIP'd from 10.000 cells and amplification of 35 pg of DNA ChIP'd from 100.000 cells (control experiment). The IP'd DNA was amplified and transformed into a sequencing-ready preparation for the Illumina plateform with the MicroPlex Library Preparation kit. The library was then analysed on an Illumina<sup>®</sup> Genome Analyzer. Cluster generation and sequencing were performed according to the manufacturer's instructions.<br />B: We observed a perfect match between the top 40% of True MicroChIP peaks and the reference dataset. Based on the NIH Encode project criterion, ChIP-seq results are considered reproducible between an original and reproduced dataset if the top 40% of peaks have at least an 80% overlap ratio with the compared dataset.</p>
<p><img src="https://www.diagenode.com/img/product/kits/true-micro-chip-result.png" alt="Cell lysis" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<p><strong>Hela cells were fixed with 1% formaldehyde (for 10 minutes at RT)</strong><br /> Cell lysis was performed using the Lysis Buffer tL1 of the Diagenode True MicroChIP kit. Samples corresponding to 10 000 cells are sheared during 5 rounds of 5 cycles of 30 seconds “ON” / 30 seconds “OFF” with the Bioruptor<sup>®</sup> Plus combined with the Bioruptor<sup>®</sup> Water cooler (Cat No. BioAcc-cool) at HIGH power setting (position H). For optimal results, samples are vortexed before and after performing 5 sonication cycles, followed by a short centrifugation at 4°C. 10 μl of DNA (equivalent to 60 000 cells) are analysed on a 1.5% agarose gel.</p>',
'label3' => '',
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'price_USD' => '590',
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'meta_title' => 'Auto True MicroChIP kit',
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'name' => 'µChIPmentation Kit for Histones',
'description' => '<p><a href="https://www.diagenode.com/files/products/kits/microchipmentation-for-histones.pdf"><img src="https://www.diagenode.com/img/buttons/bt-manual.png" /></a></p>
<p>The Diagenode <strong>µChIPmentation Kit for Histones</strong> is optimized to perform ChIP-seq on as little as <strong>10.000 cells</strong> from cell fixation to purified libraries. To <strong>reduce DNA lost</strong> the number of sample transfer from tube to tube has been limited: during whole workflow - from cell fixation to library prurification - only 3 tubes per sample are used. Reduced number of steps, reduced number of sample transfer and <a href="https://www.diagenode.com/en/categories/chromatin-ip-chipmentation" target="_blank">ChIPmentation technology</a> itself enable for <strong>efficient and robust ChIP-seq</strong> on <strong>limited amount of sample</strong>. The kit µChIPmentation for Histones includes all reagents for chromatin preparation, chromatin immunoprecipitation and library preparation using tagmentation. The primer indexes for multiplexing are not included in the kit and have to be purchase separately - <a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set1">read more</a>.</p>
<div class="extra-spaced">
<h3>Benefits of the µChIPmentation system for histone ChIP-seq</h3>
<ul>
<li><strong>Easier</strong> and <strong>faster</strong> than classical ChIP-seq</li>
<li>Optimized for <strong>low input samples</strong>: as little as 10,000 cells</li>
<li>Protocol optimized for <strong>FACS-sorted cells</strong></li>
<li>Validated for various <strong>histone marks</strong></li>
<li><strong>High quality</strong> sequencing data</li>
</ul>
</div>',
'label1' => 'Characteristics',
'info1' => '<p>ChIPmentation is based on tagmentation that allows library preparation to be integrated during the ChIP itself using transposase and sequencing-compatible adaptors. Our new improved µChIPmentation protocol combines 3 features for guaranteeing high quality sequencing data on small sample inputs 1) optimized chromatin shearing preparation protocol, 2) reduced number of steps, 3) reduced number of sample transfer from tube to tube - only 3 tubes per sample for the whole process, from cell fixation to purified libraries.</p>
<ul class="accordion" data-accordion="" style="margin-left: 0;">
<li class="accordion-navigation"><a style="background-color: white;" href="#workflow"><i class="fa fa-caret-right"></i> Workflow of µChIPmentation for Histones - Read more</a>
<div id="workflow" class="content">
<div class="extra-spaced"><center><img src="https://www.diagenode.com/img/product/kits/workflow-micro-chipmentation.png" /></center></div>
</div>
</li>
</ul>
<div class="row extra-spaced">
<div class="small-12 medium-12 large-12 columns"><strong>A.</strong><img src="https://www.diagenode.com/img/product/kits/micro-chipmentation-fig1A.png" /></div>
</div>
<div class="row extra-spaced">
<div class="small-12 medium-5 large-5 columns" columns=""><strong>B.</strong><img src="https://www.diagenode.com/img/product/kits/micro-chipmentation-fig1B.png" /></div>
<div class="small-12 medium-7 large-7 columns">
<p style="text-align: left;"><strong>Figure 1. Comparison between ChIPmentation and µChIPmentation </strong><br />ChIPmentation: chromatin preparation has been performed on 7 M K562 cells using the ChIPmentation Kit for Histones (Cat. No. C01011000) and diluted chromatin from 500.000 cells was used for the immunoprecipitation. µChIPmentation: chromatin preparation and immunoprecipitation have been performed on 10.000 K562 cells using the µChIPmentation Kit for Histones (Cat. No. C01011011) and 24 SI for Tagmented libraries (Cat. No. C010111032). The Diagenode antibody targeting H3K27me3 (Cat. No. C15410195) was used.<br /> A. Distribution of the ChIPmentation and µChIPmentation readsets in a representative region of the genome (in duplicates). B. Comparison of the top 40% peaks from µChIPmentation (10.000 cells) with dataset generated with ChIPmentation (500.000 cells).</p>
</div>
</div>
<div class="extra-spaced">
<div class="row">
<div class="small-12 medium-12 large-12 columns"><img src="https://www.diagenode.com/img/product/kits/micro-chipmentation-fig2.png" /></div>
</div>
<div class="row">
<div class="small-12 medium-12 large-12 columns">
<p><strong>Figure 2. Sequencing profiles of µChIPmentation libraries </strong><br />Chromatin preparation and immunoprecipitation have been performed on 10.000 cells using the µChIPmentation Kit for Histones (Cat. No. C01011011) and 24 SI for Tagmented libraries (Cat. No. C010111032) using K562 cells. The Diagenode antibodies targeting H3K4me3 (Cat. No. C15410003), H3K27ac (Cat. No. C15410196), H3K27me3 (Cat. No. C15410195) and H3K9me3 (Cat. No. C15410193) have been used.</p>
</div>
</div>
</div>
<div class="extra-spaced">
<div class="row">
<div class="small-12 medium-12 large-12 columns"><img src="https://www.diagenode.com/img/product/kits/micro-chipmentation-fig3.png" /></div>
</div>
<div class="row">
<div class="small-12 medium-12 large-12 columns">
<p><strong>Figure 3. Integrative genomics viewer (IGV) visualization of from ChIP-seq experiments using H3K27me3 antibody (Diagenode, Cat. No. <span>C15410195</span>) and 10.000 cells of K562 cells per immunoprecipitation.<br /></strong>Cells were FACS-sorted and ChIP has been performed accordingly to µChIPmentation protocol. Batch chromatin preparation followed by immunoprecipitation has been used per comparison as indicated.</p>
</div>
</div>
</div>',
'label2' => 'Additional solutions for µChIPmentation Kit for Histones',
'info2' => '<p><a href="https://www.diagenode.com/en/p/chromatin-shearing-optimization-kit-high-sds-100-million-cells">Chromatin EasyShear Kit - High SDS</a> optimizes chromatin shearing, a critical step for ChIP.</p>
<p><a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies" target="_blank">ChIP-seq grade anti-histone antibodies</a> provide high yields with excellent specificity and sensitivity.</p>
<p>For fast and efficient isolation of magnetic beads we recommend the magnetic racks <a href="https://www.diagenode.com/en/p/diamag02-magnetic-rack-1-unit">DiaMag0.2</a>.</p>
<p>Primer indexes for multiplexing:</p>
<ul class="no-bullet">
<li><a href="https://www.diagenode.com/en/p/24-si-for-tagmented-libraries" target="_blank">24 SI for Tagmented libraries Cat. No. C01011032</a></li>
<li><a href="https://www.diagenode.com/en/p/8-si-for-tagmented-libraries" target="_blank">8 SI for Tagmented libraries Cat. No. C01011033</a></li>
<li><em></em><a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set1">24 UDI for Tagmented libraries - Set I, Cat. No. C0101134</a></li>
<li><a href="https://www.diagenode.com/en/p/8-unique-dual-indexes-for-tagmented-libraries">8 UDI for Tagmented libraries, Cat. No. C0101135</a></li>
<li><a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set2">24 UDI for Tagmented libraries - Set II, Cat. No. C0101136</a></li>
<li><a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set3">24 UDI for Tagmented libraries - Set III, Cat. No. C0101137</a></li>
<li></li>
</ul>',
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'description' => '<div class="row">
<div class="small-12 medium-12 large-12 columns">The most important steps for a successful ChIP include both cell fixation and lysis, and chromatin shearing. Researchers often overlook the critical nature of both of these steps. Eliminating inconsistencies in the shearing step, <strong>Diagenode's Bioruptor</strong><sup>®</sup> uses state-of-the-art ultrasound <strong>ACT</strong> (<strong>A</strong>daptive <strong>C</strong>avitation <strong>T</strong>echnology) to efficiently shear chromatin. ACT enables the highest chromatin quality for high IP efficiency and sensitivity for ChIP experiments with gentle yet highly effective shearing forces. Additionally, the Bioruptor<sup>®</sup> provides a precisely controlled temperature environment that preserves chromatin from heat degradation such that protein-DNA complexes are well-preserved for sensitive, unbiased, and accurate ChIP.<br /><br /> <strong>Diagenode's Bioruptor</strong><sup>®</sup> is the instrument of choice for chromatin shearing used for a number of downstream applications such as qPCR and ChIP-seq that require optimally sheared, unbiased chromatin.</div>
<div class="small-12 medium-12 large-12 columns text-center"><br /><img src="https://www.diagenode.com/img/applications/pico_dna_shearing_fig2.png" /></div>
<div class="small-10 medium-10 large-10 columns end small-offset-1"><small> <br /><strong>Panel A, 10 µl volume:</strong> Chromatin samples are sheared for 10, 20 and 30 cycles of 30 sec ON/30 sec OFF with the Bioruptor® Pico using 0.1 ml Bioruptor® Microtubes (Cat. No. B01200041). <strong>Panel B, 100 µl volume:</strong> Chromatin samples are sheared for 10 cycles of 30 sec ON/30 sec OFF with the Bioruptor® Pico using 0.65 ml Bioruptor® Microtubes (Cat. No. WA-005-0500). <strong>Panel C, 300 µl volume:</strong> Chromatin samples are sheared for 5, 10 and 15 cycles of 30 sec ON/30 sec OFF with the Bioruptor® Pico using using 1.5 ml Bioruptor microtubes (Cat. No. C30010016). Prior to de-crosslinking, samples are treated with RNase cocktail mixture at 37°C during 1 hour. The sheared chromatin is then de-crosslinked overnight and phenol/chloroform purified as described in the kit manual. 10 µl of DNA (equivalent of 500, 000 cells) are analyzed on a 2% agarose gel (MW corresponds to the 100 bp DNA molecular weight marker).</small></div>
<div class="small-12 medium-12 large-12 columns"><br /><br /></div>
<div class="small-12 medium-12 large-12 columns">
<p>It is important to establish optimal conditions to shear crosslinked chromatin to get the correct fragment sizes needed for ChIP. Usually this process requires both optimizing sonication conditions as well as optimizing SDS concentration, which is laborious. With the Chromatin Shearing Optimization Kits, optimization is fast and easy - we provide optimization reagents with varying concentrations of SDS. Moreover, our Chromatin Shearing Optimization Kits can be used for the optimization of chromatin preparation with our kits for ChIP.</p>
</div>
<div class="small-12 medium-12 large-12 columns">
<div class="page" title="Page 7">
<table>
<tbody>
<tr valign="middle">
<td></td>
<td style="text-align: center;"><strong><a href="https://www.diagenode.com/p/chromatin-shearing-optimization-kit-low-sds-100-million-cells">Chromatin Shearing Kit Low SDS (for Histone)</a></strong></td>
<td style="text-align: center;"><strong><a href="https://www.diagenode.com/p/chromatin-shearing-optimization-kit-low-sds-for-tfs-25-rxns">Chromatin Shearing Kit Low SDS (for TF)</a></strong></td>
<td style="text-align: center;"><strong><a href="https://www.diagenode.com/p/chromatin-shearing-optimization-kit-high-sds-100-million-cells">Chromatin Shearing Kit High SDS</a></strong></td>
<td style="text-align: center;"><strong><a href="https://www.diagenode.com/p/chromatin-shearing-optimization-kit-medium-sds-100-million-cells">Chromatin Shearing Kit (for Plant)</a></strong></td>
</tr>
<tr valign="middle" style="background-color: #fff;">
<td>
<p><strong>SDS concentration</strong></p>
</td>
<td style="text-align: center;">
<p>< 0.1%</p>
</td>
<td style="text-align: center;">
<p>0.2%</p>
</td>
<td style="text-align: center;">
<p>1%</p>
</td>
<td style="text-align: center;">
<p>0.5%</p>
</td>
</tr>
<tr valign="middle" style="background-color: #fff;">
<td>
<p><strong>Nuclei isolation</strong></p>
</td>
<td style="text-align: center;">
<p>Yes</p>
</td>
<td style="text-align: center;">
<p>Yes</p>
</td>
<td style="text-align: center;">
<p>No</p>
</td>
<td style="text-align: center;">
<p>Yes</p>
</td>
</tr>
<tr valign="middle" style="background-color: #fff;">
<td>
<p><strong>Allows for shearing of... cells/tissue</strong></p>
</td>
<td style="text-align: center;">
<p>100 million cells</p>
</td>
<td style="text-align: center;">
<p>100 million cells</p>
</td>
<td style="text-align: center;">
<p>100 million cells</p>
</td>
<td style="text-align: center;">
<p>up to 25 g of tissue</p>
</td>
</tr>
<tr valign="middle" style="background-color: #fff;">
<td>
<p><strong>Corresponding to shearing buffers from</strong></p>
</td>
<td style="text-align: center;">
<p><a href="https://www.diagenode.com/en/p/ideal-chip-seq-kit-x24-24-rxns">iDeal ChIP-seq kit for Histones</a></p>
</td>
<td style="text-align: center;">
<p><a href="https://www.diagenode.com/en/p/ideal-chip-seq-kit-for-transcription-factors-x24-24-rxns">iDeal ChIP-seq Kit for Transcription Factors</a></p>
<p><a href="https://www.diagenode.com/en/p/ideal-chip-qpcr-kit">iDeal ChIP qPCR kit</a></p>
</td>
<td style="text-align: center;">
<p><a href="https://www.diagenode.com/en/p/true-microchip-kit-x16-16-rxns">True MicroChIP kit</a></p>
</td>
<td style="text-align: center;">
<p><a href="https://www.diagenode.com/en/p/universal-plant-chip-seq-kit-x24-24-rxns">Universal Plant ChIP-seq kit</a></p>
</td>
</tr>
</tbody>
</table>
<p><em><span style="font-weight: 400;">Table comes from our </span><a href="https://www.diagenode.com/protocols/bioruptor-pico-chromatin-preparation-guide"><span style="font-weight: 400;">Guide for successful chromatin preparation using the Bioruptor® Pico</span></a></em></p>
</div>
</div>
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'meta_description' => 'Diagenode's Bioruptor® is the instrument of choice for chromatin shearing used for a number of downstream applications such as qPCR and ChIP-seq that require optimally sheared, unbiased chromatin.',
'meta_title' => 'Chromatin shearing using Bioruptor® sonication device | Diagenode',
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'description' => '<p>Optimal detergent concentration is required for best results with chromatin shearing. Diagenode offers four kits with different SDS concentrations that have been pre-optimized for your specific workflow requirements. Our Chromatin EasyShear Kits (previous name: Chromatin Shearing Optimization Kits) together with the Bioruptor combine efficient cell lysis and chromatin shearing leading to consistent results.</p>
<p>The Chromatin EasyShear Kits are recommended for:</p>
<ul>
<li>The optimization of the chromatin shearing and/or chromatin preparation prior to any ChIP protocol</li>
<li>The optimization of the chromatin shearing of a new cell line/new sample type prior to ChIP using Diagenode’s ChIP kits</li>
</ul>
<p>Check all <a href="https://www.diagenode.com/en/categories/chromatin-shearing">Chromatin EasyShear Kits</a>.</p>
<p>Guide for the optimal chromatin preparation using Chromatin EasyShear Kits – <a href="https://www.diagenode.com/en/pages/chromatin-prep-easyshear-kit-guide">Read more</a></p>
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<div class="page" title="Page 4">
<div class="section">
<div class="layoutArea">
<div class="column">
<p><span>The first critical step of a successful Chromatin immunoprecipitation (ChIP) experiment is the preparation of sheared chromatin. We therefore suggest to use one of our optimized shearing ChIP kits.</span></p>
<span>• Chromatin EasyShear Kit - Ultra Low SDS <br />• Chromatin <span>EasyShear Kit</span> <span>-</span> Low SDS <br />• Chromatin EasyShear Kit - High SDS </span></div>
<div class="column"><span>• Chromatin EasyShear Kit for Plant</span><span></span>
<p><span></span></p>
<p><span>Our Chromatin EasyShear kits are used in combination with the Bioruptor</span><span>® </span><span>in order to ensure a highly reproducible chromatin shearing and make sure you obtain the right fragment size needed for your experiment. Establish optimal conditions required for shearing cross-linked chromatin (protein-DNA) is usually laborious; the protocol of the Chromatin EasyShear kits is fast, easy-to-use and optimized to get the best results possible. </span></p>
<p><span>Learn more about Diagenode <a href="https://www.diagenode.com/en/categories/chromatin-shearing">Chromatin EasyShear Kits</a>.</span></p>
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<a href="/en/p/true-microchip-kit-x16-16-rxns"><img src="/img/product/kits/chip-kit-icon.png" alt="ChIP kit icon" class="th"/></a> </div>
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<p>The Diagenode <strong>µChIPmentation Kit for Histones</strong> is optimized to perform ChIP-seq on as little as <strong>10.000 cells</strong> from cell fixation to purified libraries. To <strong>reduce DNA lost</strong> the number of sample transfer from tube to tube has been limited: during whole workflow - from cell fixation to library prurification - only 3 tubes per sample are used. Reduced number of steps, reduced number of sample transfer and <a href="https://www.diagenode.com/en/categories/chromatin-ip-chipmentation" target="_blank">ChIPmentation technology</a> itself enable for <strong>efficient and robust ChIP-seq</strong> on <strong>limited amount of sample</strong>. The kit µChIPmentation for Histones includes all reagents for chromatin preparation, chromatin immunoprecipitation and library preparation using tagmentation. The primer indexes for multiplexing are not included in the kit and have to be purchase separately - <a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set1">read more</a>.</p>
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<ul class="accordion" data-accordion="" style="margin-left: 0;">
<li class="accordion-navigation"><a style="background-color: white;" href="#workflow"><i class="fa fa-caret-right"></i> Workflow of µChIPmentation for Histones - Read more</a>
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<div class="row extra-spaced">
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<p style="text-align: left;"><strong>Figure 1. Comparison between ChIPmentation and µChIPmentation </strong><br />ChIPmentation: chromatin preparation has been performed on 7 M K562 cells using the ChIPmentation Kit for Histones (Cat. No. C01011000) and diluted chromatin from 500.000 cells was used for the immunoprecipitation. µChIPmentation: chromatin preparation and immunoprecipitation have been performed on 10.000 K562 cells using the µChIPmentation Kit for Histones (Cat. No. C01011011) and 24 SI for Tagmented libraries (Cat. No. C010111032). The Diagenode antibody targeting H3K27me3 (Cat. No. C15410195) was used.<br /> A. Distribution of the ChIPmentation and µChIPmentation readsets in a representative region of the genome (in duplicates). B. Comparison of the top 40% peaks from µChIPmentation (10.000 cells) with dataset generated with ChIPmentation (500.000 cells).</p>
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<div class="row">
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<p><strong>Figure 2. Sequencing profiles of µChIPmentation libraries </strong><br />Chromatin preparation and immunoprecipitation have been performed on 10.000 cells using the µChIPmentation Kit for Histones (Cat. No. C01011011) and 24 SI for Tagmented libraries (Cat. No. C010111032) using K562 cells. The Diagenode antibodies targeting H3K4me3 (Cat. No. C15410003), H3K27ac (Cat. No. C15410196), H3K27me3 (Cat. No. C15410195) and H3K9me3 (Cat. No. C15410193) have been used.</p>
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<p><strong>Figure 3. Integrative genomics viewer (IGV) visualization of from ChIP-seq experiments using H3K27me3 antibody (Diagenode, Cat. No. <span>C15410195</span>) and 10.000 cells of K562 cells per immunoprecipitation.<br /></strong>Cells were FACS-sorted and ChIP has been performed accordingly to µChIPmentation protocol. Batch chromatin preparation followed by immunoprecipitation has been used per comparison as indicated.</p>
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<p><a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies" target="_blank">ChIP-seq grade anti-histone antibodies</a> provide high yields with excellent specificity and sensitivity.</p>
<p>For fast and efficient isolation of magnetic beads we recommend the magnetic racks <a href="https://www.diagenode.com/en/p/diamag02-magnetic-rack-1-unit">DiaMag0.2</a>.</p>
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<li><a href="https://www.diagenode.com/en/p/8-unique-dual-indexes-for-tagmented-libraries">8 UDI for Tagmented libraries, Cat. No. C0101135</a></li>
<li><a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set2">24 UDI for Tagmented libraries - Set II, Cat. No. C0101136</a></li>
<li><a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set3">24 UDI for Tagmented libraries - Set III, Cat. No. C0101137</a></li>
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'name' => 'Chromatin shearing - Guide for successful chromatin preparation using the Bioruptor<sup>®</sup> Pico',
'description' => '<p><span>Chromatin immunoprecipitation (ChIP) is a common tool used to study protein-DNA interactions in cells and tissues. The assay can be cumbersome and the success is highly dependent on the quality of chromatin. The first critical step of a successful ChIP experiment is the preparation of sheared chromatin which is representative of the biological scenario of interest. </span></p>',
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'type' => 'Protocol',
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'name' => 'Chromatin EasyShear Kit - High SDS ',
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<p><span>The first critical step of a successful Chromatin immunoprecipitation (ChIP) experiment is the preparation of sheared chromatin. We therefore suggest to use one of our optimized shearing ChIP kits.</span></p>
<span>• Chromatin EasyShear Kit - Ultra Low SDS <br />• Chromatin <span>EasyShear Kit</span> <span>-</span> Low SDS <br />• Chromatin EasyShear Kit - High SDS </span></div>
<div class="column"><span>• Chromatin EasyShear Kit for Plant</span><span></span>
<p><span></span></p>
<p><span>Our Chromatin EasyShear kits are used in combination with the Bioruptor</span><span>® </span><span>in order to ensure a highly reproducible chromatin shearing and make sure you obtain the right fragment size needed for your experiment. Establish optimal conditions required for shearing cross-linked chromatin (protein-DNA) is usually laborious; the protocol of the Chromatin EasyShear kits is fast, easy-to-use and optimized to get the best results possible. </span></p>
<p><span>Learn more about Diagenode <a href="https://www.diagenode.com/en/categories/chromatin-shearing">Chromatin EasyShear Kits</a>.</span></p>
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'type' => 'Manual',
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'meta_keywords' => '',
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'modified' => '2021-02-01 11:51:17',
'created' => '2015-07-07 11:47:43',
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'id' => '925',
'name' => 'Chromatin EasyShear Kit - HighSDS SDS FR fr',
'language' => 'fr',
'url' => 'files/SDS/Chromatin_EasyShear_Kit-High_SDS/SDS-C01020012-Chromatin_EasyShear_Kit_-HighSDS-FR-fr-1_0.pdf',
'countries' => 'FR',
'modified' => '2020-09-23 11:31:51',
'created' => '2020-09-23 11:31:51',
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)include - APP/View/Products/view.ctp, line 755
View::_evaluate() - CORE/Cake/View/View.php, line 971
View::_render() - CORE/Cake/View/View.php, line 933
View::render() - CORE/Cake/View/View.php, line 473
Controller::render() - CORE/Cake/Controller/Controller.php, line 963
ProductsController::slug() - APP/Controller/ProductsController.php, line 1052
ReflectionMethod::invokeArgs() - [internal], line ??
Controller::invokeAction() - CORE/Cake/Controller/Controller.php, line 491
Dispatcher::_invoke() - CORE/Cake/Routing/Dispatcher.php, line 193
Dispatcher::dispatch() - CORE/Cake/Routing/Dispatcher.php, line 167
[main] - APP/webroot/index.php, line 118
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