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<h1 class="advertising-feature"><strong>Premium RRBS technology: cost-effective DNA methylation mapping with superior coverage</strong></h1>
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<li style="text-align: left;" class="vcard last-author"><a href="#auth-5" class="name"><span class="fn">Christoph Bock</span></a><sup><a href="#a2">2</a></sup><sup href="#affil-auth">, </sup></li>
<li style="text-align: left;" class="vcard"><a href="#auth-4" class="name"><span class="fn">Sharon Squazzo</span></a><sup><a href="#a3">3</a></sup><sup>, </sup></li>
<li style="text-align: left;" class="vcard"><a href="#auth-3" class="name"><span class="fn">Miklos Laczik</span></a><sup><a href="#a1">1</a></sup><sup>, </sup></li>
<li style="text-align: left;" class="vcard"><a href="#auth-2" class="name"><span class="fn">Paul Datlinger</span></a><sup><a href="#a2">2</a></sup><sup>, </sup></li>
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<p>Nature Methods 13 (2016)</p>
<p>Published online <time datetime="2016-01-28">28 January 2016 </time></p>
<p></p>
<p><a href="http://www.nature.com/nmeth/journal/v13/n2/full/nmeth.f.391.html" class="alert button"><span style="font-weight: 400;">CHECK OUT THE NATURE METHODS PAPER</span></a></p>',
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<li><strong>Cost-efficient</strong> – Multiplex up to 6 samples/sequencing lane</li>
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<p> </p>
<div class="row">
<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/kits/rrbs-figure-1.png" alt="Chr Shearing" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-6 columns"><br />
<p><b>Superior coverage</b></p>
<p>Comparison of CpG coverage between competing technologies.</p>
<p><strong><em><small>They love it! </small></em></strong><br /><em><small>The new Diagenode Premium RRBS Kit makes it easy to use RRBS cost-effectively and with high throughput, using early sample pooling and multiplex sequencing. Most importantly, the method provides an improved coverage of up to 4 million CpGs for the human genome. We successfully used this protocol on more than 1,000 samples comprising of six different species, various cancers, FFPE and lowinput samples. <br /><strong>Paul Datlinger and Christoph Bock, </strong><strong>CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria </strong></small></em></p>
<p><em> </em></p>
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<div class="row">
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<p><img src="https://www.diagenode.com/img/product/kits/rrbs-figure-2.jpg" alt="dna methylation" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<p><b>Accurate determination of DNA methylation level</b></p>
Excellent results were obtained using Diagenode’s Premium RRBS kit: almost 90% alignment rate, 4.1 million CpGs covered and bisulfite conversion rates around 99.5% for all samples. DNA methylation percentages in the region of IGF2 obtained with the Diagenode’s Premium RRBS Kit. Two human cell lines were analyzed: Gm12878 and MCF7. The MCF7 cell line was studied in duplicates. Each peak represents the DNA methylation percentage at one CpG. The methylated CpGs are shown in red and the unmethylated CpGs in grey.
<p><br /><br /></p>
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<p><img src="https://www.diagenode.com/img/landing-pages/rrbs_how_it_works.jpg" alt="rrbs how it works" style="display: block; margin-left: auto; margin-right: auto;" /><br /><br /></p>
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<div class="small-6 columns">
<p><b>How it works</b></p>
By cutting the genome using the <strong>restriction enzyme MspI</strong> (CCGG target sites) followed by size selection, DNA is enriched to represent <strong>CpG-rich genomic regions</strong> (including CpG islands, CpG island shores, enhancers, and other gene-regulatory elements), which are particularly relevant for epigenetic regulation. Similar to exome-sequencing for mutation discovery, the RRBS protocol enriches for some of the most interesting target regions and thereby achieves a reduction in sequencing cost of a factor of 10-20 compared to whole genome bisulfite sequencing.</div>
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<table width="1039">
<tbody>
<tr></tr>
<tr>
<td width="281"> </td>
<td width="379">
<h4>Premium RRBS Kit</h4>
</td>
<td width="379">
<h5>Illumina® RRBS protocol</h5>
</td>
</tr>
<tr>
<td><strong>DNA input</strong></td>
<td>100 ng</td>
<td>2-5 µg</td>
</tr>
<tr>
<td><strong>Multiplexing</strong></td>
<td>Pooling of 6 samples (one HiSeq lane)</td>
<td>no guidelines</td>
</tr>
<tr>
<td><strong>Bisulfite conversion</strong></td>
<td>One bisulfite reaction per pool (6 samples)</td>
<td>One bisulfite reaction per sample</td>
</tr>
<tr>
<td><strong># purification steps</strong></td>
<td>2</td>
<td>7</td>
</tr>
<tr>
<td><strong>Key features</strong></td>
<td width="379">low input - cost-effective - optimized for high-throughput - complete kit</td>
<td width="379">high input - not optimized for high-throughput - reagents from different providers</td>
</tr>
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<p></p>
<h1 class="advertising-feature"><strong>Premium RRBS technology: cost-effective DNA methylation mapping with superior coverage</strong></h1>
<ul class="citation dates">
<li style="text-align: left;" class="vcard last-author"><a href="#auth-5" class="name"><span class="fn">Christoph Bock</span></a><sup><a href="#a2">2</a></sup><sup href="#affil-auth">, </sup></li>
<li style="text-align: left;" class="vcard"><a href="#auth-4" class="name"><span class="fn">Sharon Squazzo</span></a><sup><a href="#a3">3</a></sup><sup>, </sup></li>
<li style="text-align: left;" class="vcard"><a href="#auth-3" class="name"><span class="fn">Miklos Laczik</span></a><sup><a href="#a1">1</a></sup><sup>, </sup></li>
<li style="text-align: left;" class="vcard"><a href="#auth-2" class="name"><span class="fn">Paul Datlinger</span></a><sup><a href="#a2">2</a></sup><sup>, </sup></li>
<li style="text-align: left;" class="vcard c1"><a href="#auth-1" class="name"><span class="fn">Anne-Clémence Veillard</span></a><sup><a href="#a1">1</a></sup><sup>, </sup></li>
</ul>
<p>Nature Methods 13 (2016)</p>
<p>Published online <time datetime="2016-01-28">28 January 2016 </time></p>
<p></p>
<p><a href="http://www.nature.com/nmeth/journal/v13/n2/full/nmeth.f.391.html" class="alert button"><span style="font-weight: 400;">CHECK OUT THE NATURE METHODS PAPER</span></a></p>
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<p>メチル化アレイとWGBS技術の長所を有するReduced representation bisulfite sequencing (RRBS) 技術は、あらゆる脊椎動物種で柔軟に使用できるため、大規模コホートの研究においても正確で費用対効果の高いアッセイを行えます。制限MspI酵素（CCGGターゲットサイト）を使用してゲノムを切断し、次にサイズを選択することで、濃縮されたDNAはプロモーターやCpGアイランドを含む関連するターゲットCpGリッチ領域を表します。またRRBS技術はゲノムの一部分を濃縮してメチル化解析を行うことによってシーケンス量を減らす手法でカバレッジが非常に高く、CpGメチル化解析に適しています。（ヒトサンプルで最大700万CpGが検出）。</p>
<h2>Genome-scale DNA methylation analysis</h2>
<ul class="square">
<li>Robust and cost-effective solution with reliable results</li>
<li>High coverage - up to 7 million CpGs detected in human samples</li>
<li>Single nucleotide resolution</li>
<li>Detection of methylation patterns in CpG rich regions including promoters and CpG islands</li>
<li>Suitable for any vertebrate species</li>
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<h2>Complete end-to-end service</h2>
<ul class="square">
<li>From DNA QC to standard bioinformatics </li>
<li>Leveraging our <a href="https://www.diagenode.com/en/p/premium-rrbs-kit-V2-x24">Premium RRBS kit V2 </a>widely adopted</li>
<li>As little as 25 ng for any vertebrate species</li>
<li>Differentially methylated site analysis </li>
</ul>
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<div class="extra-spaced">
<h2></h2>
</div>
<p><span><i class="fa fa-arrow-circle-right"></i> </span><a href="https://www.diagenode.com/en/categories/dna-methylation-profiling-services">See our other DNA Methylation Profiling Services</a></p>
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<table style="width: 905px;">
<tbody>
<tr>
<td style="width: 264px;"><strong>QC of the genomic DNA</strong></td>
<td style="width: 636px;">
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<li>Measurement of DNA concentration </li>
<li>Assessment of DNA quality</li>
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<li>Library preparation (ends preparation, adaptor ligation)</li>
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<td style="width: 264px;"><strong>Deep sequencing</strong></td>
<td style="width: 636px;">
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<li>Samples are sequenced on an Illumina platform, paired-end reads, read length 50 bp (PE50)</li>
<li>40 million raw reads (on average) per sample when pooling 10 samples/lane</li>
<li>7 million CpGs (on average) for human samples</li>
<li>7-11x CpG coverage (on average) for human samples</li>
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<li>Summary statistics</li>
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<li>Methylation level analysis</li>
<li>Differentially Methylated CpGs (DMCs) analysis</li>
<li>Differentially Methylated Regions (DMRs) analysis</li>
<li>Annotation of DMCs and DMRs for genomic regions (exons, introns, …)</li>
<li>Clustering analysis</li>
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<p><strong>Gene ontology terms analysis</strong></p>
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<li>Enrichment analysis on gene associated with DMCs and DMRs</li>
<li>Get functional insights</li>
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<p><strong>Pathway analysis</strong></p>
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<div style="text-align: justify;" class="large-12 columns">Bisulfite modification of DNA is the most commonly used, "<strong>gold standard</strong>" method for DNA methylation studies providing <strong>single nucleotide resolution</strong>. T<span style="font-weight: 400;">his technology is based on the chemical conversion of unmethylated cytosine to uracil. Methylated cytosines are protected from this conversion allowing to determine DNA methylation at the singe nucleotide level.</span></div>
<div style="text-align: justify;" class="large-12 columns"></div>
<div style="text-align: justify;" class="large-12 columns">Various analyses can be performed on the altered sequence to retrieve this information: bisulfite sequencing, pyrosequencing, methylation-specific PCR, high resolution melting curve analysis, microarray-based approaches, and next-generation sequencing.
<h3>How it works</h3>
Treatment of DNA with bisulfite converts cytosine residues to uracil, but leaves 5-methylcytosine residues unaffected (see Figure 1).
<p class="text-center"><img src="https://www.diagenode.com/img/applications/bisulfite.png" /><br />Figure 1: Overview of bisulfite conversion of DNA</p>
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<h2>Complete solutions for DNA methylation studies</h2>
<p>Whether you are experienced or new to the field of DNA methylation, Diagenode has everything you need to make your assay as easy and convenient as possible while ensuring consistent data between samples and experiments. Diagenode offers sonication instruments, reagent kits, high quality antibodies, and high-throughput automation capability to address all of your specific DNA methylation analysis requirements.</p>
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<p>DNA methylation was the first discovered epigenetic mark and is the most widely studied topic in epigenetics. <em>In vivo</em>, DNA is methylated following DNA replication and is involved in a number of biological processes including the regulation of imprinted genes, X chromosome inactivation. and tumor suppressor gene silencing in cancer cells. Methylation often occurs in cytosine-guanine rich regions of DNA (CpG islands), which are commonly upstream of promoter regions.</p>
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<div class="small-12 medium-12 large-12 columns"><br /><br />
<ul class="accordion" data-accordion="">
<li class="accordion-navigation"><a href="#dnamethyl"><i class="fa fa-caret-right"></i> Learn more</a>
<div id="dnamethyl" class="content">5-methylcytosine (5-mC) has been known for a long time as the only modification of DNA for epigenetic regulation. In 2009, however, Kriaucionis discovered a second methylated cytosine, 5-hydroxymethylcytosine (5-hmC). The so-called 6th base, is generated by enzymatic conversion of 5-methylcytosine (5-mC) into 5-hydroxymethylcytosine by the TET family of oxygenases. Early reports suggested that 5-hmC may represent an intermediate of active demethylation in a new pathway which demethylates DNA, converting 5-mC to cytosine. Recent evidence fuel this hypothesis suggesting that further oxidation of the hydroxymethyl group leads to a formyl or carboxyl group followed by either deformylation or decarboxylation. The formyl and carboxyl groups of 5-formylcytosine (5-fC) and 5-carboxylcytosine (5-caC) could be enzymatically removed without excision of the base.
<p class="text-center"><img src="https://www.diagenode.com/img/categories/kits_dna/dna_methylation_variants.jpg" /></p>
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</li>
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<br />
<h2>Main DNA methylation technologies</h2>
<p style="text-align: justify;">Overview of the <span style="font-weight: 400;">three main approaches for studying DNA methylation.</span></p>
<div class="row">
<ol>
<li style="font-weight: 400;"><span style="font-weight: 400;">Chemical modification with bisulfite – Bisulfite conversion</span></li>
<li style="font-weight: 400;"><span style="font-weight: 400;">Enrichment of methylated DNA (including MeDIP and MBD)</span></li>
<li style="font-weight: 400;"><span style="font-weight: 400;">Treatment with methylation-sensitive or dependent restriction enzymes</span></li>
</ol>
<p><span style="font-weight: 400;"> </span></p>
<div class="row">
<table>
<thead>
<tr>
<th></th>
<th>Description</th>
<th width="350">Features</th>
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</thead>
<tbody>
<tr>
<td><strong>Bisulfite conversion</strong></td>
<td><span style="font-weight: 400;">Chemical conversion of unmethylated cytosine to uracil. Methylated cytosines are protected from this conversion allowing to determine DNA methylation at single nucleotide resolution.</span></td>
<td>
<ul style="list-style-type: circle;">
<li style="font-weight: 400;"><span style="font-weight: 400;">Single nucleotide resolution</span></li>
<li style="font-weight: 400;"><span style="font-weight: 400;">Quantitative analysis - methylation rate (%)</span></li>
<li style="font-weight: 400;"><span style="font-weight: 400;">Gold standard and well studied</span></li>
<li style="font-weight: 400;"><span style="font-weight: 400;">Compatible with automation</span></li>
</ul>
</td>
</tr>
<tr>
<td><b>Methylated DNA enrichment</b></td>
<td><span style="font-weight: 400;">(Hydroxy-)Methylated DNA is enriched by using specific antibodies (hMeDIP or MeDIP) or proteins (MBD) that specifically bind methylated CpG sites in fragmented genomic DNA.</span></td>
<td>
<ul style="list-style-type: circle;">
<li style="font-weight: 400;"><span style="font-weight: 400;">Resolution depends on the fragment size of the enriched methylated DNA (300 bp)</span></li>
<li style="font-weight: 400;"><span style="font-weight: 400;">Qualitative analysis</span></li>
<li style="font-weight: 400;"><span style="font-weight: 400;">Compatible with automation</span></li>
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</td>
</tr>
<tr>
<td><strong>Restriction enzyme-based digestion</strong></td>
<td><span style="font-weight: 400;">Use of (hydroxy)methylation-sensitive or (hydroxy)methylation-dependent restriction enzymes for DNA methylation analysis at specific sites.</span></td>
<td>
<ul style="list-style-type: circle;">
<li style="font-weight: 400;"><span style="font-weight: 400;">Determination of methylation status is limited by the enzyme recognition site</span></li>
<li style="font-weight: 400;"><span style="font-weight: 400;">Easy to use</span></li>
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<p>Sodium bisulfite conversion of genomic DNA is the most commonly used method for DNA methylation studies providing <strong>single nucleotide resolution</strong>. It enables <span>to differentiate and detect unmethylated versus methylated cytosines. This procedure can then be followed either by <strong>PCR amplification</strong> or <strong>next generation sequencing</strong> to reveal the methylation status of every cytosine in gene specific amplification or whole genome amplification.</span></p>
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<h2>How it works</h2>
<p style="text-align: left;">Treatment of DNA with sodium bisulfite converts unmethylated cytosine to uracil, while methylated cytosines remain unchanged. <span>The DNA is then amplified by PCR where the uracils are converted to thymines. </span></p>
<p style="text-align: center;"><span></span></p>
<p><img src="https://www.diagenode.com/img/categories/bisulfite-conversion/bisulfite-conversion-acgautac.png" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<h2>Advantages</h2>
<ul class="nobullet" style="font-size: 19px;">
<li><i class="fa fa-arrow-circle-right"></i><strong> </strong><strong>Single nucleotide</strong> resolution</li>
<li><i class="fa fa-arrow-circle-right"></i><strong> Gene-specific </strong>and <strong>genome-wide</strong><span> analyses</span></li>
<li><i class="fa fa-arrow-circle-right"></i><strong> NGS</strong><span> </span>compatible</li>
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<h2>Downstream analysis techniques</h2>
<ul class="square">
<li>Reduced Representation Bisulfite Sequencing (RRBS) with our <a href="https://www.diagenode.com/en/p/premium-rrbs-kit-V2-x24">Premium RRBS Kit V2</a></li>
<li>Bisulfite conversion with our <a href="https://www.diagenode.com/en/p/premium-bisulfite-kit-50-rxns">Premium Bisulfite Kit</a> followed by qPCR, Sanger, Pyrosequencing</li>
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<p><span>Positive and negative spike-in controls are included for the monitoring of bisulfite conversion efficiency. </span></p>
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<li>
<p><span>Size selection has been optimized to keep small fragments of interest and to remove adaptor dimers, resulting in a better coverag. </span></p>
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<p><span>The pooling protocol includes a quantification of the samples and a pooling application, available on our website, to help you to choose the groups, depending on the DNA amount and adaptor barcode of each sample. </span></p>
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<p><span>The bisulfite conversion protocol has been improved to decrease DNA degradation while keeping a highly efficient conversion of unmethylated cytosine. </span></p>
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<div class="row">
<div class="small-12 columns">
<a href="/en/p/premium-rrbs-kit-x24-24-rxns"><img src="/img/product/kits/methyl-kit-icon.png" alt="Methylation kit icon" class="th"/></a> </div>
<div class="small-12 columns">
<div class="small-6 columns" style="padding-left:0px;padding-right:0px;margin-top:-6px;margin-left:-1px">
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<!--a href="#" style="color:#B21329"><i class="fa fa-info-circle"></i></a-->
</div>
</div>
<div class="small-12 columns" >
<h6 style="height:60px">Premium RRBS kit (24 rxns) - replaced by Premiu...</h6>
</div>
</div>
</li>
<li>
<div class="row">
<div class="small-12 columns">
<a href="/en/p/premium-rrbs-kit-x96-96-rxns"><img src="/img/product/kits/methyl-kit-icon.png" alt="Methylation kit icon" class="th"/></a> </div>
<div class="small-12 columns">
<div class="small-6 columns" style="padding-left:0px;padding-right:0px;margin-top:-6px;margin-left:-1px">
<span class="success label" style="">C02030033</span>
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<div class="small-6 columns text-right" style="padding-left:0px;padding-right:0px;margin-top:-6px">
<!--a href="#" style="color:#B21329"><i class="fa fa-info-circle"></i></a-->
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</div>
<div class="small-12 columns" >
<h6 style="height:60px">Premium RRBS kit (96 rxns) - replaced by Premiu...</h6>
</div>
</div>
</li>
<li>
<div class="row">
<div class="small-12 columns">
<a href="/en/p/rrbs-service"><img src="/img/grey-logo.jpg" alt="default alt" class="th"/></a> </div>
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<div class="row">
<div class="small-12 medium-12 large-12 columns">
<h3>Get a quote</h3><p class="lead">You are about to request a quote for <strong>our epigenomics services</strong>. Fill out the form below and we will be in touch with you very soon.</p><p><small>All <span style="font-size:16px;color:red;">*</span> fields are mandatory</small></p>
</div>
</div>
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<h2>Service Information</h2>
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<div class="small-12 large-12 columns">
<h4>Which services are you interested in?</h4>
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<div class="small-12 large-12 columns">
<input type="hidden" name="data[Quote][epigenomics_service]" value="" id="QuoteEpigenomicsService"/>
<div class="checkbox"><input type="checkbox" name="data[Quote][epigenomics_service][]" value="ChIP-seq" id="QuoteEpigenomicsServiceChIPSeq" /><label for="QuoteEpigenomicsServiceChIPSeq">ChIP-seq</label></div>
<div class="checkbox"><input type="checkbox" name="data[Quote][epigenomics_service][]" value="ATAC-seq" id="QuoteEpigenomicsServiceATACSeq" /><label for="QuoteEpigenomicsServiceATACSeq">ATAC-seq</label></div>
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<div class="checkbox"><input type="checkbox" name="data[Quote][epigenomics_service][]" value="WGBS" id="QuoteEpigenomicsServiceWGBS" /><label for="QuoteEpigenomicsServiceWGBS">WGBS</label></div>
<div class="checkbox"><input type="checkbox" name="data[Quote][epigenomics_service][]" value="MeDIP-seq" id="QuoteEpigenomicsServiceMeDIPSeq" /><label for="QuoteEpigenomicsServiceMeDIPSeq">MeDIP-seq</label></div>
<div class="checkbox"><input type="checkbox" name="data[Quote][epigenomics_service][]" value="Targeted DNA methylation analysis" id="QuoteEpigenomicsServiceTargetedDNAMethylationAnalysis" /><label for="QuoteEpigenomicsServiceTargetedDNAMethylationAnalysis">Targeted DNA methylation analysis</label></div>
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<div class="checkbox"><input type="checkbox" name="data[Quote][epigenomics_service][]" value="Human Methylome" id="QuoteEpigenomicsServiceHumanMethylome" /><label for="QuoteEpigenomicsServiceHumanMethylome">Human Methylome</label></div>
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<div class="small-12 medium-12 large-3 columns">
<span class="prefix">Sample species</span>
</div>
<div class="small-12 medium-12 large-9 columns">
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<div class="row collapse">
<div class="small-12 medium-12 large-6 columns">
<span class="prefix">Total number of samples (including replicates)</span>
</div>
<div class="small-12 medium-12 large-6 columns">
<input name="data[Quote][number_samples]" maxlength="255" type="text" id="QuoteNumberSamples"/> </div>
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<h2>Contact Information</h2>
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<span class="prefix">First name <sup style="font-size:16px;color:red;">*</sup></span>
</div>
<div class="small-9 large-10 columns">
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<span class="prefix">Company <sup style="font-size:16px;color:red;">*</sup></span>
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<input name="data[Quote][company]" placeholder="Organisation / Institute" maxlength="255" type="text" id="QuoteCompany" required="required"/> </div>
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<span class="prefix">Phone number</span>
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<option value="AL">Albania</option>
<option value="DZ">Algeria</option>
<option value="AS">American Samoa</option>
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<option value="AQ">Antarctica</option>
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<option value="CX">Christmas Island</option>
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<option value="HM">Heard Island and McDonald Islands</option>
<option value="VA">Holy See (Vatican City State)</option>
<option value="HN">Honduras</option>
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<p>メチル化アレイとWGBS技術の長所を有するReduced representation bisulfite sequencing (RRBS) 技術は、あらゆる脊椎動物種で柔軟に使用できるため、大規模コホートの研究においても正確で費用対効果の高いアッセイを行えます。制限MspI酵素（CCGGターゲットサイト）を使用してゲノムを切断し、次にサイズを選択することで、濃縮されたDNAはプロモーターやCpGアイランドを含む関連するターゲットCpGリッチ領域を表します。またRRBS技術はゲノムの一部分を濃縮してメチル化解析を行うことによってシーケンス量を減らす手法でカバレッジが非常に高く、CpGメチル化解析に適しています。（ヒトサンプルで最大700万CpGが検出）。</p>
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<h2>Complete end-to-end service</h2>
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<li>Differentially methylated site analysis </li>
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<h2></h2>
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<p><span><i class="fa fa-arrow-circle-right"></i> </span><a href="https://www.diagenode.com/en/categories/dna-methylation-profiling-services">See our other DNA Methylation Profiling Services</a></p>
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<p><strong>Gene ontology terms analysis</strong></p>
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<li>Get functional insights</li>
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<h1 class="advertising-feature"><strong>Premium RRBS technology: cost-effective DNA methylation mapping with superior coverage</strong></h1>
<ul class="citation dates">
<li style="text-align: left;" class="vcard last-author"><a href="#auth-5" class="name"><span class="fn">Christoph Bock</span></a><sup><a href="#a2">2</a></sup><sup href="#affil-auth">, </sup></li>
<li style="text-align: left;" class="vcard"><a href="#auth-4" class="name"><span class="fn">Sharon Squazzo</span></a><sup><a href="#a3">3</a></sup><sup>, </sup></li>
<li style="text-align: left;" class="vcard"><a href="#auth-3" class="name"><span class="fn">Miklos Laczik</span></a><sup><a href="#a1">1</a></sup><sup>, </sup></li>
<li style="text-align: left;" class="vcard"><a href="#auth-2" class="name"><span class="fn">Paul Datlinger</span></a><sup><a href="#a2">2</a></sup><sup>, </sup></li>
<li style="text-align: left;" class="vcard c1"><a href="#auth-1" class="name"><span class="fn">Anne-Clémence Veillard</span></a><sup><a href="#a1">1</a></sup><sup>, </sup></li>
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<p>Nature Methods 13 (2016)</p>
<p>Published online <time datetime="2016-01-28">28 January 2016 </time></p>
<p></p>
<p><a href="http://www.nature.com/nmeth/journal/v13/n2/full/nmeth.f.391.html" class="alert button"><span style="font-weight: 400;">CHECK OUT THE NATURE METHODS PAPER</span></a></p>',
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<li><strong>Cost-efficient</strong> – Multiplex up to 6 samples/sequencing lane</li>
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<li class="accordion-navigation" style="list-style-type: circle; display: list-item;"><strong>Already tested on various species </strong>– human, mouse, rat, pig, cow, dog, zebrafish, Daphnia <a href="#species" style="color: #13b29c; background-color: transparent; display: inline; padding: 0;">and more</a> <span class="content" id="species">cichlid fish (Astatotilapia calliptera), mossy frog, yellow-bellied slider, dice snake, zebra finch, humboldt penguin, leaf bird, buzzard, vulturine guinea fowe, parma kangaroo, cheetah, mouflon</span></li>
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<div class="small-6 columns"><br />
<p><b>Superior coverage</b></p>
<p>Comparison of CpG coverage between competing technologies.</p>
<p><strong><em><small>They love it! </small></em></strong><br /><em><small>The new Diagenode Premium RRBS Kit makes it easy to use RRBS cost-effectively and with high throughput, using early sample pooling and multiplex sequencing. Most importantly, the method provides an improved coverage of up to 4 million CpGs for the human genome. We successfully used this protocol on more than 1,000 samples comprising of six different species, various cancers, FFPE and lowinput samples. <br /><strong>Paul Datlinger and Christoph Bock, </strong><strong>CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria </strong></small></em></p>
<p><em> </em></p>
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<div class="row">
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<div class="small-6 columns">
<p><b>Accurate determination of DNA methylation level</b></p>
Excellent results were obtained using Diagenode’s Premium RRBS kit: almost 90% alignment rate, 4.1 million CpGs covered and bisulfite conversion rates around 99.5% for all samples. DNA methylation percentages in the region of IGF2 obtained with the Diagenode’s Premium RRBS Kit. Two human cell lines were analyzed: Gm12878 and MCF7. The MCF7 cell line was studied in duplicates. Each peak represents the DNA methylation percentage at one CpG. The methylated CpGs are shown in red and the unmethylated CpGs in grey.
<p><br /><br /></p>
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<p><img src="https://www.diagenode.com/img/landing-pages/rrbs_how_it_works.jpg" alt="rrbs how it works" style="display: block; margin-left: auto; margin-right: auto;" /><br /><br /></p>
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<div class="small-6 columns">
<p><b>How it works</b></p>
By cutting the genome using the <strong>restriction enzyme MspI</strong> (CCGG target sites) followed by size selection, DNA is enriched to represent <strong>CpG-rich genomic regions</strong> (including CpG islands, CpG island shores, enhancers, and other gene-regulatory elements), which are particularly relevant for epigenetic regulation. Similar to exome-sequencing for mutation discovery, the RRBS protocol enriches for some of the most interesting target regions and thereby achieves a reduction in sequencing cost of a factor of 10-20 compared to whole genome bisulfite sequencing.</div>
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<table width="1039">
<tbody>
<tr></tr>
<tr>
<td width="281"> </td>
<td width="379">
<h4>Premium RRBS Kit</h4>
</td>
<td width="379">
<h5>Illumina® RRBS protocol</h5>
</td>
</tr>
<tr>
<td><strong>DNA input</strong></td>
<td>100 ng</td>
<td>2-5 µg</td>
</tr>
<tr>
<td><strong>Multiplexing</strong></td>
<td>Pooling of 6 samples (one HiSeq lane)</td>
<td>no guidelines</td>
</tr>
<tr>
<td><strong>Bisulfite conversion</strong></td>
<td>One bisulfite reaction per pool (6 samples)</td>
<td>One bisulfite reaction per sample</td>
</tr>
<tr>
<td><strong># purification steps</strong></td>
<td>2</td>
<td>7</td>
</tr>
<tr>
<td><strong>Key features</strong></td>
<td width="379">low input - cost-effective - optimized for high-throughput - complete kit</td>
<td width="379">high input - not optimized for high-throughput - reagents from different providers</td>
</tr>
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<h1 class="advertising-feature"><strong>Premium RRBS technology: cost-effective DNA methylation mapping with superior coverage</strong></h1>
<ul class="citation dates">
<li style="text-align: left;" class="vcard last-author"><a href="#auth-5" class="name"><span class="fn">Christoph Bock</span></a><sup><a href="#a2">2</a></sup><sup href="#affil-auth">, </sup></li>
<li style="text-align: left;" class="vcard"><a href="#auth-4" class="name"><span class="fn">Sharon Squazzo</span></a><sup><a href="#a3">3</a></sup><sup>, </sup></li>
<li style="text-align: left;" class="vcard"><a href="#auth-3" class="name"><span class="fn">Miklos Laczik</span></a><sup><a href="#a1">1</a></sup><sup>, </sup></li>
<li style="text-align: left;" class="vcard"><a href="#auth-2" class="name"><span class="fn">Paul Datlinger</span></a><sup><a href="#a2">2</a></sup><sup>, </sup></li>
<li style="text-align: left;" class="vcard c1"><a href="#auth-1" class="name"><span class="fn">Anne-Clémence Veillard</span></a><sup><a href="#a1">1</a></sup><sup>, </sup></li>
</ul>
<p>Nature Methods 13 (2016)</p>
<p>Published online <time datetime="2016-01-28">28 January 2016 </time></p>
<p></p>
<p><a href="http://www.nature.com/nmeth/journal/v13/n2/full/nmeth.f.391.html" class="alert button"><span style="font-weight: 400;">CHECK OUT THE NATURE METHODS PAPER</span></a></p>
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<p>メチル化アレイとWGBS技術の長所を有するReduced representation bisulfite sequencing (RRBS) 技術は、あらゆる脊椎動物種で柔軟に使用できるため、大規模コホートの研究においても正確で費用対効果の高いアッセイを行えます。制限MspI酵素（CCGGターゲットサイト）を使用してゲノムを切断し、次にサイズを選択することで、濃縮されたDNAはプロモーターやCpGアイランドを含む関連するターゲットCpGリッチ領域を表します。またRRBS技術はゲノムの一部分を濃縮してメチル化解析を行うことによってシーケンス量を減らす手法でカバレッジが非常に高く、CpGメチル化解析に適しています。（ヒトサンプルで最大700万CpGが検出）。</p>
<h2>Genome-scale DNA methylation analysis</h2>
<ul class="square">
<li>Robust and cost-effective solution with reliable results</li>
<li>High coverage - up to 7 million CpGs detected in human samples</li>
<li>Single nucleotide resolution</li>
<li>Detection of methylation patterns in CpG rich regions including promoters and CpG islands</li>
<li>Suitable for any vertebrate species</li>
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<h2>Complete end-to-end service</h2>
<ul class="square">
<li>From DNA QC to standard bioinformatics </li>
<li>Leveraging our <a href="https://www.diagenode.com/en/p/premium-rrbs-kit-V2-x24">Premium RRBS kit V2 </a>widely adopted</li>
<li>As little as 25 ng for any vertebrate species</li>
<li>Differentially methylated site analysis </li>
</ul>
</div>
<div class="extra-spaced">
<h2></h2>
</div>
<p><span><i class="fa fa-arrow-circle-right"></i> </span><a href="https://www.diagenode.com/en/categories/dna-methylation-profiling-services">See our other DNA Methylation Profiling Services</a></p>
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<tr>
<td style="width: 264px;"><strong>QC of the genomic DNA</strong></td>
<td style="width: 636px;">
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<li>Measurement of DNA concentration </li>
<li>Assessment of DNA quality</li>
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<td style="width: 264px;"><strong>Preparation of RRBS libraries</strong></td>
<td style="width: 636px;">
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<li>Size selection</li>
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<li>Library amplification and clean-up</li>
<li>QC of the RRBS library pool (DNA concentration, analysis of the pool profile)</li>
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<tr>
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<td style="width: 636px;">
<ul style="list-style-type: circle;">
<li>Samples are sequenced on an Illumina platform, paired-end reads, read length 50 bp (PE50)</li>
<li>40 million raw reads (on average) per sample when pooling 10 samples/lane</li>
<li>7 million CpGs (on average) for human samples</li>
<li>7-11x CpG coverage (on average) for human samples</li>
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<h4><strong>Analysis</strong></h4>
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<h4><strong>Features</strong></h4>
</th>
</tr>
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<td style="width: 262px;"><strong>Standard</strong></td>
<td style="width: 624px;">
<ul>
<li>FASTQ raw data</li>
<li>FASTQC quality control insights</li>
<li>Alignment of bisulfite sequencing data against reference genome</li>
<li>Methylation calling and extraction</li>
<li>Summary statistics</li>
</ul>
</td>
</tr>
<tr>
<td style="width: 262px;"><strong>Differential methylation analysis<br /></strong></td>
<td style="width: 624px;">
<ul>
<li>Methylation level analysis</li>
<li>Differentially Methylated CpGs (DMCs) analysis</li>
<li>Differentially Methylated Regions (DMRs) analysis</li>
<li>Annotation of DMCs and DMRs for genomic regions (exons, introns, …)</li>
<li>Clustering analysis</li>
</ul>
</td>
</tr>
<tr>
<td style="width: 262px;">
<p><strong>Gene ontology terms analysis</strong></p>
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<td style="width: 624px;">
<ul>
<li>Enrichment analysis on gene associated with DMCs and DMRs</li>
<li>Get functional insights</li>
</ul>
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</tr>
<tr>
<td style="width: 262px;">
<p><strong>Pathway analysis</strong></p>
</td>
<td style="width: 624px;">
<ul>
<li>Identification of biological pathways in which genes associated with DMCs and DMRs may be over-represented (or under-represented)</li>
<li>Get mechanistic insights</li>
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<div style="text-align: justify;" class="large-12 columns">Bisulfite modification of DNA is the most commonly used, "<strong>gold standard</strong>" method for DNA methylation studies providing <strong>single nucleotide resolution</strong>. T<span style="font-weight: 400;">his technology is based on the chemical conversion of unmethylated cytosine to uracil. Methylated cytosines are protected from this conversion allowing to determine DNA methylation at the singe nucleotide level.</span></div>
<div style="text-align: justify;" class="large-12 columns"></div>
<div style="text-align: justify;" class="large-12 columns">Various analyses can be performed on the altered sequence to retrieve this information: bisulfite sequencing, pyrosequencing, methylation-specific PCR, high resolution melting curve analysis, microarray-based approaches, and next-generation sequencing.
<h3>How it works</h3>
Treatment of DNA with bisulfite converts cytosine residues to uracil, but leaves 5-methylcytosine residues unaffected (see Figure 1).
<p class="text-center"><img src="https://www.diagenode.com/img/applications/bisulfite.png" /><br />Figure 1: Overview of bisulfite conversion of DNA</p>
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<h2>Complete solutions for DNA methylation studies</h2>
<p>Whether you are experienced or new to the field of DNA methylation, Diagenode has everything you need to make your assay as easy and convenient as possible while ensuring consistent data between samples and experiments. Diagenode offers sonication instruments, reagent kits, high quality antibodies, and high-throughput automation capability to address all of your specific DNA methylation analysis requirements.</p>
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<p>DNA methylation was the first discovered epigenetic mark and is the most widely studied topic in epigenetics. <em>In vivo</em>, DNA is methylated following DNA replication and is involved in a number of biological processes including the regulation of imprinted genes, X chromosome inactivation. and tumor suppressor gene silencing in cancer cells. Methylation often occurs in cytosine-guanine rich regions of DNA (CpG islands), which are commonly upstream of promoter regions.</p>
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<div class="small-12 medium-12 large-12 columns"><br /><br />
<ul class="accordion" data-accordion="">
<li class="accordion-navigation"><a href="#dnamethyl"><i class="fa fa-caret-right"></i> Learn more</a>
<div id="dnamethyl" class="content">5-methylcytosine (5-mC) has been known for a long time as the only modification of DNA for epigenetic regulation. In 2009, however, Kriaucionis discovered a second methylated cytosine, 5-hydroxymethylcytosine (5-hmC). The so-called 6th base, is generated by enzymatic conversion of 5-methylcytosine (5-mC) into 5-hydroxymethylcytosine by the TET family of oxygenases. Early reports suggested that 5-hmC may represent an intermediate of active demethylation in a new pathway which demethylates DNA, converting 5-mC to cytosine. Recent evidence fuel this hypothesis suggesting that further oxidation of the hydroxymethyl group leads to a formyl or carboxyl group followed by either deformylation or decarboxylation. The formyl and carboxyl groups of 5-formylcytosine (5-fC) and 5-carboxylcytosine (5-caC) could be enzymatically removed without excision of the base.
<p class="text-center"><img src="https://www.diagenode.com/img/categories/kits_dna/dna_methylation_variants.jpg" /></p>
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</li>
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<br />
<h2>Main DNA methylation technologies</h2>
<p style="text-align: justify;">Overview of the <span style="font-weight: 400;">three main approaches for studying DNA methylation.</span></p>
<div class="row">
<ol>
<li style="font-weight: 400;"><span style="font-weight: 400;">Chemical modification with bisulfite – Bisulfite conversion</span></li>
<li style="font-weight: 400;"><span style="font-weight: 400;">Enrichment of methylated DNA (including MeDIP and MBD)</span></li>
<li style="font-weight: 400;"><span style="font-weight: 400;">Treatment with methylation-sensitive or dependent restriction enzymes</span></li>
</ol>
<p><span style="font-weight: 400;"> </span></p>
<div class="row">
<table>
<thead>
<tr>
<th></th>
<th>Description</th>
<th width="350">Features</th>
</tr>
</thead>
<tbody>
<tr>
<td><strong>Bisulfite conversion</strong></td>
<td><span style="font-weight: 400;">Chemical conversion of unmethylated cytosine to uracil. Methylated cytosines are protected from this conversion allowing to determine DNA methylation at single nucleotide resolution.</span></td>
<td>
<ul style="list-style-type: circle;">
<li style="font-weight: 400;"><span style="font-weight: 400;">Single nucleotide resolution</span></li>
<li style="font-weight: 400;"><span style="font-weight: 400;">Quantitative analysis - methylation rate (%)</span></li>
<li style="font-weight: 400;"><span style="font-weight: 400;">Gold standard and well studied</span></li>
<li style="font-weight: 400;"><span style="font-weight: 400;">Compatible with automation</span></li>
</ul>
</td>
</tr>
<tr>
<td><b>Methylated DNA enrichment</b></td>
<td><span style="font-weight: 400;">(Hydroxy-)Methylated DNA is enriched by using specific antibodies (hMeDIP or MeDIP) or proteins (MBD) that specifically bind methylated CpG sites in fragmented genomic DNA.</span></td>
<td>
<ul style="list-style-type: circle;">
<li style="font-weight: 400;"><span style="font-weight: 400;">Resolution depends on the fragment size of the enriched methylated DNA (300 bp)</span></li>
<li style="font-weight: 400;"><span style="font-weight: 400;">Qualitative analysis</span></li>
<li style="font-weight: 400;"><span style="font-weight: 400;">Compatible with automation</span></li>
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</td>
</tr>
<tr>
<td><strong>Restriction enzyme-based digestion</strong></td>
<td><span style="font-weight: 400;">Use of (hydroxy)methylation-sensitive or (hydroxy)methylation-dependent restriction enzymes for DNA methylation analysis at specific sites.</span></td>
<td>
<ul style="list-style-type: circle;">
<li style="font-weight: 400;"><span style="font-weight: 400;">Determination of methylation status is limited by the enzyme recognition site</span></li>
<li style="font-weight: 400;"><span style="font-weight: 400;">Easy to use</span></li>
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<p>Sodium bisulfite conversion of genomic DNA is the most commonly used method for DNA methylation studies providing <strong>single nucleotide resolution</strong>. It enables <span>to differentiate and detect unmethylated versus methylated cytosines. This procedure can then be followed either by <strong>PCR amplification</strong> or <strong>next generation sequencing</strong> to reveal the methylation status of every cytosine in gene specific amplification or whole genome amplification.</span></p>
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<h2>How it works</h2>
<p style="text-align: left;">Treatment of DNA with sodium bisulfite converts unmethylated cytosine to uracil, while methylated cytosines remain unchanged. <span>The DNA is then amplified by PCR where the uracils are converted to thymines. </span></p>
<p style="text-align: center;"><span></span></p>
<p><img src="https://www.diagenode.com/img/categories/bisulfite-conversion/bisulfite-conversion-acgautac.png" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<h2>Advantages</h2>
<ul class="nobullet" style="font-size: 19px;">
<li><i class="fa fa-arrow-circle-right"></i><strong> </strong><strong>Single nucleotide</strong> resolution</li>
<li><i class="fa fa-arrow-circle-right"></i><strong> Gene-specific </strong>and <strong>genome-wide</strong><span> analyses</span></li>
<li><i class="fa fa-arrow-circle-right"></i><strong> NGS</strong><span> </span>compatible</li>
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<h2>Downstream analysis techniques</h2>
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<li>Reduced Representation Bisulfite Sequencing (RRBS) with our <a href="https://www.diagenode.com/en/p/premium-rrbs-kit-V2-x24">Premium RRBS Kit V2</a></li>
<li>Bisulfite conversion with our <a href="https://www.diagenode.com/en/p/premium-bisulfite-kit-50-rxns">Premium Bisulfite Kit</a> followed by qPCR, Sanger, Pyrosequencing</li>
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<p><span>Positive and negative spike-in controls are included for the monitoring of bisulfite conversion efficiency. </span></p>
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<p><span>Size selection has been optimized to keep small fragments of interest and to remove adaptor dimers, resulting in a better coverag. </span></p>
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<h3>Get a quote</h3><p class="lead">You are about to request a quote for <strong>our epigenomics services</strong>. Fill out the form below and we will be in touch with you very soon.</p><p><small>All <span style="font-size:16px;color:red;">*</span> fields are mandatory</small></p>
</div>
</div>
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<h2>Service Information</h2>
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<div class="small-12 large-12 columns">
<h4>Which services are you interested in?</h4>
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<div class="small-12 large-12 columns">
<input type="hidden" name="data[Quote][epigenomics_service]" value="" id="QuoteEpigenomicsService"/>
<div class="checkbox"><input type="checkbox" name="data[Quote][epigenomics_service][]" value="ChIP-seq" id="QuoteEpigenomicsServiceChIPSeq" /><label for="QuoteEpigenomicsServiceChIPSeq">ChIP-seq</label></div>
<div class="checkbox"><input type="checkbox" name="data[Quote][epigenomics_service][]" value="ATAC-seq" id="QuoteEpigenomicsServiceATACSeq" /><label for="QuoteEpigenomicsServiceATACSeq">ATAC-seq</label></div>
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<div class="checkbox"><input type="checkbox" name="data[Quote][epigenomics_service][]" value="WGBS" id="QuoteEpigenomicsServiceWGBS" /><label for="QuoteEpigenomicsServiceWGBS">WGBS</label></div>
<div class="checkbox"><input type="checkbox" name="data[Quote][epigenomics_service][]" value="MeDIP-seq" id="QuoteEpigenomicsServiceMeDIPSeq" /><label for="QuoteEpigenomicsServiceMeDIPSeq">MeDIP-seq</label></div>
<div class="checkbox"><input type="checkbox" name="data[Quote][epigenomics_service][]" value="Targeted DNA methylation analysis" id="QuoteEpigenomicsServiceTargetedDNAMethylationAnalysis" /><label for="QuoteEpigenomicsServiceTargetedDNAMethylationAnalysis">Targeted DNA methylation analysis</label></div>
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<div class="checkbox"><input type="checkbox" name="data[Quote][epigenomics_service][]" value="Infinium Mouse Methylation Array" id="QuoteEpigenomicsServiceInfiniumMouseMethylationArray" /><label for="QuoteEpigenomicsServiceInfiniumMouseMethylationArray">Infinium Mouse Methylation Array</label></div>
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<div class="checkbox"><input type="checkbox" name="data[Quote][epigenomics_service][]" value="Data mining" id="QuoteEpigenomicsServiceDataMining" /><label for="QuoteEpigenomicsServiceDataMining">Data mining</label></div>
<div class="checkbox"><input type="checkbox" name="data[Quote][epigenomics_service][]" value="Human Methylome" id="QuoteEpigenomicsServiceHumanMethylome" /><label for="QuoteEpigenomicsServiceHumanMethylome">Human Methylome</label></div>
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<span class="prefix">Sample species</span>
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<div class="small-12 medium-12 large-9 columns">
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<span class="prefix">Total number of samples (including replicates)</span>
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<h2>Contact Information</h2>
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<span class="prefix">First name <sup style="font-size:16px;color:red;">*</sup></span>
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<div class="small-9 large-10 columns">
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<span class="prefix">Company <sup style="font-size:16px;color:red;">*</sup></span>
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<span class="prefix">Phone number</span>
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<option value="AS">American Samoa</option>
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<option value="AO">Angola</option>
<option value="AI">Anguilla</option>
<option value="AQ">Antarctica</option>
<option value="AG">Antigua and Barbuda</option>
<option value="AR">Argentina</option>
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<option value="CV">Cape Verde</option>
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<option value="TD">Chad</option>
<option value="CL">Chile</option>
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<option value="CX">Christmas Island</option>
<option value="CC">Cocos (Keeling) Islands</option>
<option value="CO">Colombia</option>
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<option value="CG">Congo</option>
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<option value="HR">Croatia</option>
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<option value="EC">Ecuador</option>
<option value="EG">Egypt</option>
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<option value="ER">Eritrea</option>
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<h6 style="height:60px">RRBS Service (Reduced Representation Bisulfite ...</h6>
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<p>メチル化アレイとWGBS技術の長所を有するReduced representation bisulfite sequencing (RRBS) 技術は、あらゆる脊椎動物種で柔軟に使用できるため、大規模コホートの研究においても正確で費用対効果の高いアッセイを行えます。制限MspI酵素（CCGGターゲットサイト）を使用してゲノムを切断し、次にサイズを選択することで、濃縮されたDNAはプロモーターやCpGアイランドを含む関連するターゲットCpGリッチ領域を表します。またRRBS技術はゲノムの一部分を濃縮してメチル化解析を行うことによってシーケンス量を減らす手法でカバレッジが非常に高く、CpGメチル化解析に適しています。（ヒトサンプルで最大700万CpGが検出）。</p>
<h2>Genome-scale DNA methylation analysis</h2>
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<li>High coverage - up to 7 million CpGs detected in human samples</li>
<li>Single nucleotide resolution</li>
<li>Detection of methylation patterns in CpG rich regions including promoters and CpG islands</li>
<li>Suitable for any vertebrate species</li>
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<h2>Complete end-to-end service</h2>
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<li>Leveraging our <a href="https://www.diagenode.com/en/p/premium-rrbs-kit-V2-x24">Premium RRBS kit V2 </a>widely adopted</li>
<li>As little as 25 ng for any vertebrate species</li>
<li>Differentially methylated site analysis </li>
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<h2></h2>
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<p><span><i class="fa fa-arrow-circle-right"></i> </span><a href="https://www.diagenode.com/en/categories/dna-methylation-profiling-services">See our other DNA Methylation Profiling Services</a></p>
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<li>Get functional insights</li>
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<p style="text-align: justify;"><span>Reduced Representation Bisulfite Sequencing provides a powerful method to efficiently analyze DNA methylation at the </span><strong>single nucleotide level</strong><span> without the higher costs associated with whole genome bisulfite sequencing. By cutting the genome using the restriction MspI enzyme (CCGG target sites) followed by size selection, DNA is enriched to represent CpG-rich regions (including CpG islands), in which DNA methylation marks are typically found. Thus, RRBS provides a </span><strong>cost-effective</strong><span> method for analyzing DNA methylation by reducing the part of the genome that actually needs to be sequenced and focusing on relevant CpG islands.</span></p>',
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<h1 class="advertising-feature"><strong>Premium RRBS technology: cost-effective DNA methylation mapping with superior coverage</strong></h1>
<ul class="citation dates">
<li style="text-align: left;" class="vcard last-author"><a href="#auth-5" class="name"><span class="fn">Christoph Bock</span></a><sup><a href="#a2">2</a></sup><sup href="#affil-auth">, </sup></li>
<li style="text-align: left;" class="vcard"><a href="#auth-4" class="name"><span class="fn">Sharon Squazzo</span></a><sup><a href="#a3">3</a></sup><sup>, </sup></li>
<li style="text-align: left;" class="vcard"><a href="#auth-3" class="name"><span class="fn">Miklos Laczik</span></a><sup><a href="#a1">1</a></sup><sup>, </sup></li>
<li style="text-align: left;" class="vcard"><a href="#auth-2" class="name"><span class="fn">Paul Datlinger</span></a><sup><a href="#a2">2</a></sup><sup>, </sup></li>
<li style="text-align: left;" class="vcard c1"><a href="#auth-1" class="name"><span class="fn">Anne-Clémence Veillard</span></a><sup><a href="#a1">1</a></sup><sup>, </sup></li>
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<p>Nature Methods 13 (2016)</p>
<p>Published online <time datetime="2016-01-28">28 January 2016 </time></p>
<p></p>
<p><a href="http://www.nature.com/nmeth/journal/v13/n2/full/nmeth.f.391.html" class="alert button"><span style="font-weight: 400;">CHECK OUT THE NATURE METHODS PAPER</span></a></p>',
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<li><strong>Superior coverage</strong> – 4 million CpGs</li>
<li><strong>High throughput</strong> - 96 samples processed in one experiment</li>
<li><strong>Cost-efficient</strong> – Multiplex up to 6 samples/sequencing lane</li>
<li><strong>Validated</strong> with FFPE, cancer, and low-input samples</li>
<li><strong>High efficiency and minimal bias</strong> - Low DNA degradation and reduced amplification</li>
<li><strong>Complete kit</strong> – Bisulfite conversion reagents, MspI enzyme, library preparation reagents including barcodes, and spike-in controls</li>
<li class="accordion-navigation" style="list-style-type: circle; display: list-item;"><strong>Already tested on various species </strong>– human, mouse, rat, pig, cow, dog, zebrafish, Daphnia <a href="#species" style="color: #13b29c; background-color: transparent; display: inline; padding: 0;">and more</a> <span class="content" id="species">cichlid fish (Astatotilapia calliptera), mossy frog, yellow-bellied slider, dice snake, zebra finch, humboldt penguin, leaf bird, buzzard, vulturine guinea fowe, parma kangaroo, cheetah, mouflon</span></li>
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<p> </p>
<div class="row">
<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/kits/rrbs-figure-1.png" alt="Chr Shearing" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-6 columns"><br />
<p><b>Superior coverage</b></p>
<p>Comparison of CpG coverage between competing technologies.</p>
<p><strong><em><small>They love it! </small></em></strong><br /><em><small>The new Diagenode Premium RRBS Kit makes it easy to use RRBS cost-effectively and with high throughput, using early sample pooling and multiplex sequencing. Most importantly, the method provides an improved coverage of up to 4 million CpGs for the human genome. We successfully used this protocol on more than 1,000 samples comprising of six different species, various cancers, FFPE and lowinput samples. <br /><strong>Paul Datlinger and Christoph Bock, </strong><strong>CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria </strong></small></em></p>
<p><em> </em></p>
</div>
</div>
<div class="row">
<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/kits/rrbs-figure-2.jpg" alt="dna methylation" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-6 columns">
<p><b>Accurate determination of DNA methylation level</b></p>
Excellent results were obtained using Diagenode’s Premium RRBS kit: almost 90% alignment rate, 4.1 million CpGs covered and bisulfite conversion rates around 99.5% for all samples. DNA methylation percentages in the region of IGF2 obtained with the Diagenode’s Premium RRBS Kit. Two human cell lines were analyzed: Gm12878 and MCF7. The MCF7 cell line was studied in duplicates. Each peak represents the DNA methylation percentage at one CpG. The methylated CpGs are shown in red and the unmethylated CpGs in grey.
<p><br /><br /></p>
</div>
</div>
<div class="row">
<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/landing-pages/rrbs_how_it_works.jpg" alt="rrbs how it works" style="display: block; margin-left: auto; margin-right: auto;" /><br /><br /></p>
</div>
<div class="small-6 columns">
<p><b>How it works</b></p>
By cutting the genome using the <strong>restriction enzyme MspI</strong> (CCGG target sites) followed by size selection, DNA is enriched to represent <strong>CpG-rich genomic regions</strong> (including CpG islands, CpG island shores, enhancers, and other gene-regulatory elements), which are particularly relevant for epigenetic regulation. Similar to exome-sequencing for mutation discovery, the RRBS protocol enriches for some of the most interesting target regions and thereby achieves a reduction in sequencing cost of a factor of 10-20 compared to whole genome bisulfite sequencing.</div>
</div>
<table width="1039">
<tbody>
<tr></tr>
<tr>
<td width="281"> </td>
<td width="379">
<h4>Premium RRBS Kit</h4>
</td>
<td width="379">
<h5>Illumina® RRBS protocol</h5>
</td>
</tr>
<tr>
<td><strong>DNA input</strong></td>
<td>100 ng</td>
<td>2-5 µg</td>
</tr>
<tr>
<td><strong>Multiplexing</strong></td>
<td>Pooling of 6 samples (one HiSeq lane)</td>
<td>no guidelines</td>
</tr>
<tr>
<td><strong>Bisulfite conversion</strong></td>
<td>One bisulfite reaction per pool (6 samples)</td>
<td>One bisulfite reaction per sample</td>
</tr>
<tr>
<td><strong># purification steps</strong></td>
<td>2</td>
<td>7</td>
</tr>
<tr>
<td><strong>Key features</strong></td>
<td width="379">low input - cost-effective - optimized for high-throughput - complete kit</td>
<td width="379">high input - not optimized for high-throughput - reagents from different providers</td>
</tr>
</tbody>
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<p></p>
<h1 class="advertising-feature"><strong>Premium RRBS technology: cost-effective DNA methylation mapping with superior coverage</strong></h1>
<ul class="citation dates">
<li style="text-align: left;" class="vcard last-author"><a href="#auth-5" class="name"><span class="fn">Christoph Bock</span></a><sup><a href="#a2">2</a></sup><sup href="#affil-auth">, </sup></li>
<li style="text-align: left;" class="vcard"><a href="#auth-4" class="name"><span class="fn">Sharon Squazzo</span></a><sup><a href="#a3">3</a></sup><sup>, </sup></li>
<li style="text-align: left;" class="vcard"><a href="#auth-3" class="name"><span class="fn">Miklos Laczik</span></a><sup><a href="#a1">1</a></sup><sup>, </sup></li>
<li style="text-align: left;" class="vcard"><a href="#auth-2" class="name"><span class="fn">Paul Datlinger</span></a><sup><a href="#a2">2</a></sup><sup>, </sup></li>
<li style="text-align: left;" class="vcard c1"><a href="#auth-1" class="name"><span class="fn">Anne-Clémence Veillard</span></a><sup><a href="#a1">1</a></sup><sup>, </sup></li>
</ul>
<p>Nature Methods 13 (2016)</p>
<p>Published online <time datetime="2016-01-28">28 January 2016 </time></p>
<p></p>
<p><a href="http://www.nature.com/nmeth/journal/v13/n2/full/nmeth.f.391.html" class="alert button"><span style="font-weight: 400;">CHECK OUT THE NATURE METHODS PAPER</span></a></p>
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<div class="extra-spaced">
<p>メチル化アレイとWGBS技術の長所を有するReduced representation bisulfite sequencing (RRBS) 技術は、あらゆる脊椎動物種で柔軟に使用できるため、大規模コホートの研究においても正確で費用対効果の高いアッセイを行えます。制限MspI酵素（CCGGターゲットサイト）を使用してゲノムを切断し、次にサイズを選択することで、濃縮されたDNAはプロモーターやCpGアイランドを含む関連するターゲットCpGリッチ領域を表します。またRRBS技術はゲノムの一部分を濃縮してメチル化解析を行うことによってシーケンス量を減らす手法でカバレッジが非常に高く、CpGメチル化解析に適しています。（ヒトサンプルで最大700万CpGが検出）。</p>
<h2>Genome-scale DNA methylation analysis</h2>
<ul class="square">
<li>Robust and cost-effective solution with reliable results</li>
<li>High coverage - up to 7 million CpGs detected in human samples</li>
<li>Single nucleotide resolution</li>
<li>Detection of methylation patterns in CpG rich regions including promoters and CpG islands</li>
<li>Suitable for any vertebrate species</li>
</ul>
<h2>Complete end-to-end service</h2>
<ul class="square">
<li>From DNA QC to standard bioinformatics </li>
<li>Leveraging our <a href="https://www.diagenode.com/en/p/premium-rrbs-kit-V2-x24">Premium RRBS kit V2 </a>widely adopted</li>
<li>As little as 25 ng for any vertebrate species</li>
<li>Differentially methylated site analysis </li>
</ul>
</div>
<div class="extra-spaced">
<h2></h2>
</div>
<p><span><i class="fa fa-arrow-circle-right"></i> </span><a href="https://www.diagenode.com/en/categories/dna-methylation-profiling-services">See our other DNA Methylation Profiling Services</a></p>
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<h4>RRBS Service includes:</h4>
<table style="width: 905px;">
<tbody>
<tr>
<td style="width: 264px;"><strong>QC of the genomic DNA</strong></td>
<td style="width: 636px;">
<ul style="list-style-type: circle;">
<li>Measurement of DNA concentration </li>
<li>Assessment of DNA quality</li>
</ul>
</td>
</tr>
<tr>
<td style="width: 264px;"><strong>Preparation of RRBS libraries</strong></td>
<td style="width: 636px;">
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<li>MspI digestion</li>
<li>Library preparation (ends preparation, adaptor ligation)</li>
<li>Size selection</li>
<li>Sample pooling</li>
<li>Bisulfite conversion</li>
<li>Library amplification and clean-up</li>
<li>QC of the RRBS library pool (DNA concentration, analysis of the pool profile)</li>
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<tr>
<td style="width: 264px;"><strong>Deep sequencing</strong></td>
<td style="width: 636px;">
<ul style="list-style-type: circle;">
<li>Samples are sequenced on an Illumina platform, paired-end reads, read length 50 bp (PE50)</li>
<li>40 million raw reads (on average) per sample when pooling 10 samples/lane</li>
<li>7 million CpGs (on average) for human samples</li>
<li>7-11x CpG coverage (on average) for human samples</li>
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<h4><strong>Analysis</strong></h4>
</th>
<th style="width: 624px;">
<h4><strong>Features</strong></h4>
</th>
</tr>
</thead>
<tbody>
<tr>
<td style="width: 262px;"><strong>Standard</strong></td>
<td style="width: 624px;">
<ul>
<li>FASTQ raw data</li>
<li>FASTQC quality control insights</li>
<li>Alignment of bisulfite sequencing data against reference genome</li>
<li>Methylation calling and extraction</li>
<li>Summary statistics</li>
</ul>
</td>
</tr>
<tr>
<td style="width: 262px;"><strong>Differential methylation analysis<br /></strong></td>
<td style="width: 624px;">
<ul>
<li>Methylation level analysis</li>
<li>Differentially Methylated CpGs (DMCs) analysis</li>
<li>Differentially Methylated Regions (DMRs) analysis</li>
<li>Annotation of DMCs and DMRs for genomic regions (exons, introns, …)</li>
<li>Clustering analysis</li>
</ul>
</td>
</tr>
<tr>
<td style="width: 262px;">
<p><strong>Gene ontology terms analysis</strong></p>
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<td style="width: 624px;">
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<li>Enrichment analysis on gene associated with DMCs and DMRs</li>
<li>Get functional insights</li>
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<tr>
<td style="width: 262px;">
<p><strong>Pathway analysis</strong></p>
</td>
<td style="width: 624px;">
<ul>
<li>Identification of biological pathways in which genes associated with DMCs and DMRs may be over-represented (or under-represented)</li>
<li>Get mechanistic insights</li>
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<div style="text-align: justify;" class="large-12 columns">Bisulfite modification of DNA is the most commonly used, "<strong>gold standard</strong>" method for DNA methylation studies providing <strong>single nucleotide resolution</strong>. T<span style="font-weight: 400;">his technology is based on the chemical conversion of unmethylated cytosine to uracil. Methylated cytosines are protected from this conversion allowing to determine DNA methylation at the singe nucleotide level.</span></div>
<div style="text-align: justify;" class="large-12 columns"></div>
<div style="text-align: justify;" class="large-12 columns">Various analyses can be performed on the altered sequence to retrieve this information: bisulfite sequencing, pyrosequencing, methylation-specific PCR, high resolution melting curve analysis, microarray-based approaches, and next-generation sequencing.
<h3>How it works</h3>
Treatment of DNA with bisulfite converts cytosine residues to uracil, but leaves 5-methylcytosine residues unaffected (see Figure 1).
<p class="text-center"><img src="https://www.diagenode.com/img/applications/bisulfite.png" /><br />Figure 1: Overview of bisulfite conversion of DNA</p>
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<h2>Complete solutions for DNA methylation studies</h2>
<p>Whether you are experienced or new to the field of DNA methylation, Diagenode has everything you need to make your assay as easy and convenient as possible while ensuring consistent data between samples and experiments. Diagenode offers sonication instruments, reagent kits, high quality antibodies, and high-throughput automation capability to address all of your specific DNA methylation analysis requirements.</p>
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<p>DNA methylation was the first discovered epigenetic mark and is the most widely studied topic in epigenetics. <em>In vivo</em>, DNA is methylated following DNA replication and is involved in a number of biological processes including the regulation of imprinted genes, X chromosome inactivation. and tumor suppressor gene silencing in cancer cells. Methylation often occurs in cytosine-guanine rich regions of DNA (CpG islands), which are commonly upstream of promoter regions.</p>
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<div class="small-12 medium-12 large-12 columns"><br /><br />
<ul class="accordion" data-accordion="">
<li class="accordion-navigation"><a href="#dnamethyl"><i class="fa fa-caret-right"></i> Learn more</a>
<div id="dnamethyl" class="content">5-methylcytosine (5-mC) has been known for a long time as the only modification of DNA for epigenetic regulation. In 2009, however, Kriaucionis discovered a second methylated cytosine, 5-hydroxymethylcytosine (5-hmC). The so-called 6th base, is generated by enzymatic conversion of 5-methylcytosine (5-mC) into 5-hydroxymethylcytosine by the TET family of oxygenases. Early reports suggested that 5-hmC may represent an intermediate of active demethylation in a new pathway which demethylates DNA, converting 5-mC to cytosine. Recent evidence fuel this hypothesis suggesting that further oxidation of the hydroxymethyl group leads to a formyl or carboxyl group followed by either deformylation or decarboxylation. The formyl and carboxyl groups of 5-formylcytosine (5-fC) and 5-carboxylcytosine (5-caC) could be enzymatically removed without excision of the base.
<p class="text-center"><img src="https://www.diagenode.com/img/categories/kits_dna/dna_methylation_variants.jpg" /></p>
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</li>
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<br />
<h2>Main DNA methylation technologies</h2>
<p style="text-align: justify;">Overview of the <span style="font-weight: 400;">three main approaches for studying DNA methylation.</span></p>
<div class="row">
<ol>
<li style="font-weight: 400;"><span style="font-weight: 400;">Chemical modification with bisulfite – Bisulfite conversion</span></li>
<li style="font-weight: 400;"><span style="font-weight: 400;">Enrichment of methylated DNA (including MeDIP and MBD)</span></li>
<li style="font-weight: 400;"><span style="font-weight: 400;">Treatment with methylation-sensitive or dependent restriction enzymes</span></li>
</ol>
<p><span style="font-weight: 400;"> </span></p>
<div class="row">
<table>
<thead>
<tr>
<th></th>
<th>Description</th>
<th width="350">Features</th>
</tr>
</thead>
<tbody>
<tr>
<td><strong>Bisulfite conversion</strong></td>
<td><span style="font-weight: 400;">Chemical conversion of unmethylated cytosine to uracil. Methylated cytosines are protected from this conversion allowing to determine DNA methylation at single nucleotide resolution.</span></td>
<td>
<ul style="list-style-type: circle;">
<li style="font-weight: 400;"><span style="font-weight: 400;">Single nucleotide resolution</span></li>
<li style="font-weight: 400;"><span style="font-weight: 400;">Quantitative analysis - methylation rate (%)</span></li>
<li style="font-weight: 400;"><span style="font-weight: 400;">Gold standard and well studied</span></li>
<li style="font-weight: 400;"><span style="font-weight: 400;">Compatible with automation</span></li>
</ul>
</td>
</tr>
<tr>
<td><b>Methylated DNA enrichment</b></td>
<td><span style="font-weight: 400;">(Hydroxy-)Methylated DNA is enriched by using specific antibodies (hMeDIP or MeDIP) or proteins (MBD) that specifically bind methylated CpG sites in fragmented genomic DNA.</span></td>
<td>
<ul style="list-style-type: circle;">
<li style="font-weight: 400;"><span style="font-weight: 400;">Resolution depends on the fragment size of the enriched methylated DNA (300 bp)</span></li>
<li style="font-weight: 400;"><span style="font-weight: 400;">Qualitative analysis</span></li>
<li style="font-weight: 400;"><span style="font-weight: 400;">Compatible with automation</span></li>
</ul>
</td>
</tr>
<tr>
<td><strong>Restriction enzyme-based digestion</strong></td>
<td><span style="font-weight: 400;">Use of (hydroxy)methylation-sensitive or (hydroxy)methylation-dependent restriction enzymes for DNA methylation analysis at specific sites.</span></td>
<td>
<ul style="list-style-type: circle;">
<li style="font-weight: 400;"><span style="font-weight: 400;">Determination of methylation status is limited by the enzyme recognition site</span></li>
<li style="font-weight: 400;"><span style="font-weight: 400;">Easy to use</span></li>
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<p>Sodium bisulfite conversion of genomic DNA is the most commonly used method for DNA methylation studies providing <strong>single nucleotide resolution</strong>. It enables <span>to differentiate and detect unmethylated versus methylated cytosines. This procedure can then be followed either by <strong>PCR amplification</strong> or <strong>next generation sequencing</strong> to reveal the methylation status of every cytosine in gene specific amplification or whole genome amplification.</span></p>
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<h2>How it works</h2>
<p style="text-align: left;">Treatment of DNA with sodium bisulfite converts unmethylated cytosine to uracil, while methylated cytosines remain unchanged. <span>The DNA is then amplified by PCR where the uracils are converted to thymines. </span></p>
<p style="text-align: center;"><span></span></p>
<p><img src="https://www.diagenode.com/img/categories/bisulfite-conversion/bisulfite-conversion-acgautac.png" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<h2>Advantages</h2>
<ul class="nobullet" style="font-size: 19px;">
<li><i class="fa fa-arrow-circle-right"></i><strong> </strong><strong>Single nucleotide</strong> resolution</li>
<li><i class="fa fa-arrow-circle-right"></i><strong> Gene-specific </strong>and <strong>genome-wide</strong><span> analyses</span></li>
<li><i class="fa fa-arrow-circle-right"></i><strong> NGS</strong><span> </span>compatible</li>
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<h2>Downstream analysis techniques</h2>
<ul class="square">
<li>Reduced Representation Bisulfite Sequencing (RRBS) with our <a href="https://www.diagenode.com/en/p/premium-rrbs-kit-V2-x24">Premium RRBS Kit V2</a></li>
<li>Bisulfite conversion with our <a href="https://www.diagenode.com/en/p/premium-bisulfite-kit-50-rxns">Premium Bisulfite Kit</a> followed by qPCR, Sanger, Pyrosequencing</li>
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<p><span>Positive and negative spike-in controls are included for the monitoring of bisulfite conversion efficiency. </span></p>
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<p><span>Size selection has been optimized to keep small fragments of interest and to remove adaptor dimers, resulting in a better coverag. </span></p>
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<p><span>The pooling protocol includes a quantification of the samples and a pooling application, available on our website, to help you to choose the groups, depending on the DNA amount and adaptor barcode of each sample. </span></p>
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<p><span>The bisulfite conversion protocol has been improved to decrease DNA degradation while keeping a highly efficient conversion of unmethylated cytosine. </span></p>
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<h6 style="height:60px">Premium RRBS kit (24 rxns) - replaced by Premiu...</h6>
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<a href="/en/p/premium-rrbs-kit-x96-96-rxns"><img src="/img/product/kits/methyl-kit-icon.png" alt="Methylation kit icon" class="th"/></a> </div>
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<h6 style="height:60px">Premium RRBS kit (96 rxns) - replaced by Premiu...</h6>
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<h3>Get a quote</h3><p class="lead">You are about to request a quote for <strong>our epigenomics services</strong>. Fill out the form below and we will be in touch with you very soon.</p><p><small>All <span style="font-size:16px;color:red;">*</span> fields are mandatory</small></p>
</div>
</div>
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<h2>Service Information</h2>
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<div class="small-12 large-12 columns">
<h4>Which services are you interested in?</h4>
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<div class="small-12 large-12 columns">
<input type="hidden" name="data[Quote][epigenomics_service]" value="" id="QuoteEpigenomicsService"/>
<div class="checkbox"><input type="checkbox" name="data[Quote][epigenomics_service][]" value="ChIP-seq" id="QuoteEpigenomicsServiceChIPSeq" /><label for="QuoteEpigenomicsServiceChIPSeq">ChIP-seq</label></div>
<div class="checkbox"><input type="checkbox" name="data[Quote][epigenomics_service][]" value="ATAC-seq" id="QuoteEpigenomicsServiceATACSeq" /><label for="QuoteEpigenomicsServiceATACSeq">ATAC-seq</label></div>
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<div class="checkbox"><input type="checkbox" name="data[Quote][epigenomics_service][]" value="WGBS" id="QuoteEpigenomicsServiceWGBS" /><label for="QuoteEpigenomicsServiceWGBS">WGBS</label></div>
<div class="checkbox"><input type="checkbox" name="data[Quote][epigenomics_service][]" value="MeDIP-seq" id="QuoteEpigenomicsServiceMeDIPSeq" /><label for="QuoteEpigenomicsServiceMeDIPSeq">MeDIP-seq</label></div>
<div class="checkbox"><input type="checkbox" name="data[Quote][epigenomics_service][]" value="Targeted DNA methylation analysis" id="QuoteEpigenomicsServiceTargetedDNAMethylationAnalysis" /><label for="QuoteEpigenomicsServiceTargetedDNAMethylationAnalysis">Targeted DNA methylation analysis</label></div>
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<div class="checkbox"><input type="checkbox" name="data[Quote][epigenomics_service][]" value="Infinium Mouse Methylation Array" id="QuoteEpigenomicsServiceInfiniumMouseMethylationArray" /><label for="QuoteEpigenomicsServiceInfiniumMouseMethylationArray">Infinium Mouse Methylation Array</label></div>
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<div class="checkbox"><input type="checkbox" name="data[Quote][epigenomics_service][]" value="Data mining" id="QuoteEpigenomicsServiceDataMining" /><label for="QuoteEpigenomicsServiceDataMining">Data mining</label></div>
<div class="checkbox"><input type="checkbox" name="data[Quote][epigenomics_service][]" value="Human Methylome" id="QuoteEpigenomicsServiceHumanMethylome" /><label for="QuoteEpigenomicsServiceHumanMethylome">Human Methylome</label></div>
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<div class="row collapse">
<div class="small-12 medium-12 large-3 columns">
<span class="prefix">Sample species</span>
</div>
<div class="small-12 medium-12 large-9 columns">
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</div>
<div class="row collapse">
<div class="small-12 medium-12 large-6 columns">
<span class="prefix">Total number of samples (including replicates)</span>
</div>
<div class="small-12 medium-12 large-6 columns">
<input name="data[Quote][number_samples]" maxlength="255" type="text" id="QuoteNumberSamples"/> </div>
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<div class="row collapse">
<h2>Contact Information</h2>
<div class="small-3 large-2 columns">
<span class="prefix">First name <sup style="font-size:16px;color:red;">*</sup></span>
</div>
<div class="small-9 large-10 columns">
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<span class="prefix">Company <sup style="font-size:16px;color:red;">*</sup></span>
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<span class="prefix">Phone number</span>
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<div class="small-9 large-10 columns">
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<div class="small-3 large-2 columns">
<span class="prefix">City</span>
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<option value="AF">Afghanistan</option>
<option value="AX">Åland Islands</option>
<option value="AL">Albania</option>
<option value="DZ">Algeria</option>
<option value="AS">American Samoa</option>
<option value="AD">Andorra</option>
<option value="AO">Angola</option>
<option value="AI">Anguilla</option>
<option value="AQ">Antarctica</option>
<option value="AG">Antigua and Barbuda</option>
<option value="AR">Argentina</option>
<option value="AM">Armenia</option>
<option value="AW">Aruba</option>
<option value="AU">Australia</option>
<option value="AT">Austria</option>
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<option value="BW">Botswana</option>
<option value="BV">Bouvet Island</option>
<option value="BR">Brazil</option>
<option value="IO">British Indian Ocean Territory</option>
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<option value="BG">Bulgaria</option>
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<option value="BI">Burundi</option>
<option value="KH">Cambodia</option>
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<option value="CV">Cape Verde</option>
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<option value="CF">Central African Republic</option>
<option value="TD">Chad</option>
<option value="CL">Chile</option>
<option value="CN">China</option>
<option value="CX">Christmas Island</option>
<option value="CC">Cocos (Keeling) Islands</option>
<option value="CO">Colombia</option>
<option value="KM">Comoros</option>
<option value="CG">Congo</option>
<option value="CD">Congo, The Democratic Republic of the</option>
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<option value="CR">Costa Rica</option>
<option value="CI">Côte d'Ivoire</option>
<option value="HR">Croatia</option>
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<option value="CW">Curaçao</option>
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<option value="DJ">Djibouti</option>
<option value="DM">Dominica</option>
<option value="DO">Dominican Republic</option>
<option value="EC">Ecuador</option>
<option value="EG">Egypt</option>
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<option value="GQ">Equatorial Guinea</option>
<option value="ER">Eritrea</option>
<option value="EE">Estonia</option>
<option value="ET">Ethiopia</option>
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<option value="GM">Gambia</option>
<option value="GE">Georgia</option>
<option value="DE">Germany</option>
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<option value="GD">Grenada</option>
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<h6 style="height:60px">RRBS Service (Reduced Representation Bisulfite ...</h6>
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<p>メチル化アレイとWGBS技術の長所を有するReduced representation bisulfite sequencing (RRBS) 技術は、あらゆる脊椎動物種で柔軟に使用できるため、大規模コホートの研究においても正確で費用対効果の高いアッセイを行えます。制限MspI酵素（CCGGターゲットサイト）を使用してゲノムを切断し、次にサイズを選択することで、濃縮されたDNAはプロモーターやCpGアイランドを含む関連するターゲットCpGリッチ領域を表します。またRRBS技術はゲノムの一部分を濃縮してメチル化解析を行うことによってシーケンス量を減らす手法でカバレッジが非常に高く、CpGメチル化解析に適しています。（ヒトサンプルで最大700万CpGが検出）。</p>
<h2>Genome-scale DNA methylation analysis</h2>
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<li>High coverage - up to 7 million CpGs detected in human samples</li>
<li>Single nucleotide resolution</li>
<li>Detection of methylation patterns in CpG rich regions including promoters and CpG islands</li>
<li>Suitable for any vertebrate species</li>
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<h2>Complete end-to-end service</h2>
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<li>Leveraging our <a href="https://www.diagenode.com/en/p/premium-rrbs-kit-V2-x24">Premium RRBS kit V2 </a>widely adopted</li>
<li>As little as 25 ng for any vertebrate species</li>
<li>Differentially methylated site analysis </li>
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<h2></h2>
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<p><span><i class="fa fa-arrow-circle-right"></i> </span><a href="https://www.diagenode.com/en/categories/dna-methylation-profiling-services">See our other DNA Methylation Profiling Services</a></p>
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<h4><strong>Analysis</strong></h4>
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<td style="width: 262px;"><strong>Standard</strong></td>
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<li>Get functional insights</li>
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<p style="text-align: justify;"><span>Reduced Representation Bisulfite Sequencing provides a powerful method to efficiently analyze DNA methylation at the </span><strong>single nucleotide level</strong><span> without the higher costs associated with whole genome bisulfite sequencing. By cutting the genome using the restriction MspI enzyme (CCGG target sites) followed by size selection, DNA is enriched to represent CpG-rich regions (including CpG islands), in which DNA methylation marks are typically found. Thus, RRBS provides a </span><strong>cost-effective</strong><span> method for analyzing DNA methylation by reducing the part of the genome that actually needs to be sequenced and focusing on relevant CpG islands.</span></p>',
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<h1 class="advertising-feature"><strong>Premium RRBS technology: cost-effective DNA methylation mapping with superior coverage</strong></h1>
<ul class="citation dates">
<li style="text-align: left;" class="vcard last-author"><a href="#auth-5" class="name"><span class="fn">Christoph Bock</span></a><sup><a href="#a2">2</a></sup><sup href="#affil-auth">, </sup></li>
<li style="text-align: left;" class="vcard"><a href="#auth-4" class="name"><span class="fn">Sharon Squazzo</span></a><sup><a href="#a3">3</a></sup><sup>, </sup></li>
<li style="text-align: left;" class="vcard"><a href="#auth-3" class="name"><span class="fn">Miklos Laczik</span></a><sup><a href="#a1">1</a></sup><sup>, </sup></li>
<li style="text-align: left;" class="vcard"><a href="#auth-2" class="name"><span class="fn">Paul Datlinger</span></a><sup><a href="#a2">2</a></sup><sup>, </sup></li>
<li style="text-align: left;" class="vcard c1"><a href="#auth-1" class="name"><span class="fn">Anne-Clémence Veillard</span></a><sup><a href="#a1">1</a></sup><sup>, </sup></li>
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<p>Nature Methods 13 (2016)</p>
<p>Published online <time datetime="2016-01-28">28 January 2016 </time></p>
<p></p>
<p><a href="http://www.nature.com/nmeth/journal/v13/n2/full/nmeth.f.391.html" class="alert button"><span style="font-weight: 400;">CHECK OUT THE NATURE METHODS PAPER</span></a></p>',
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<li><strong>Superior coverage</strong> – 4 million CpGs</li>
<li><strong>High throughput</strong> - 96 samples processed in one experiment</li>
<li><strong>Cost-efficient</strong> – Multiplex up to 6 samples/sequencing lane</li>
<li><strong>Validated</strong> with FFPE, cancer, and low-input samples</li>
<li><strong>High efficiency and minimal bias</strong> - Low DNA degradation and reduced amplification</li>
<li><strong>Complete kit</strong> – Bisulfite conversion reagents, MspI enzyme, library preparation reagents including barcodes, and spike-in controls</li>
<li class="accordion-navigation" style="list-style-type: circle; display: list-item;"><strong>Already tested on various species </strong>– human, mouse, rat, pig, cow, dog, zebrafish, Daphnia <a href="#species" style="color: #13b29c; background-color: transparent; display: inline; padding: 0;">and more</a> <span class="content" id="species">cichlid fish (Astatotilapia calliptera), mossy frog, yellow-bellied slider, dice snake, zebra finch, humboldt penguin, leaf bird, buzzard, vulturine guinea fowe, parma kangaroo, cheetah, mouflon</span></li>
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<p> </p>
<div class="row">
<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/kits/rrbs-figure-1.png" alt="Chr Shearing" style="display: block; margin-left: auto; margin-right: auto;" /></p>
</div>
<div class="small-6 columns"><br />
<p><b>Superior coverage</b></p>
<p>Comparison of CpG coverage between competing technologies.</p>
<p><strong><em><small>They love it! </small></em></strong><br /><em><small>The new Diagenode Premium RRBS Kit makes it easy to use RRBS cost-effectively and with high throughput, using early sample pooling and multiplex sequencing. Most importantly, the method provides an improved coverage of up to 4 million CpGs for the human genome. We successfully used this protocol on more than 1,000 samples comprising of six different species, various cancers, FFPE and lowinput samples. <br /><strong>Paul Datlinger and Christoph Bock, </strong><strong>CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria </strong></small></em></p>
<p><em> </em></p>
</div>
</div>
<div class="row">
<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/product/kits/rrbs-figure-2.jpg" alt="dna methylation" style="display: block; margin-left: auto; margin-right: auto;" /></p>
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<div class="small-6 columns">
<p><b>Accurate determination of DNA methylation level</b></p>
Excellent results were obtained using Diagenode’s Premium RRBS kit: almost 90% alignment rate, 4.1 million CpGs covered and bisulfite conversion rates around 99.5% for all samples. DNA methylation percentages in the region of IGF2 obtained with the Diagenode’s Premium RRBS Kit. Two human cell lines were analyzed: Gm12878 and MCF7. The MCF7 cell line was studied in duplicates. Each peak represents the DNA methylation percentage at one CpG. The methylated CpGs are shown in red and the unmethylated CpGs in grey.
<p><br /><br /></p>
</div>
</div>
<div class="row">
<div class="small-6 columns">
<p><img src="https://www.diagenode.com/img/landing-pages/rrbs_how_it_works.jpg" alt="rrbs how it works" style="display: block; margin-left: auto; margin-right: auto;" /><br /><br /></p>
</div>
<div class="small-6 columns">
<p><b>How it works</b></p>
By cutting the genome using the <strong>restriction enzyme MspI</strong> (CCGG target sites) followed by size selection, DNA is enriched to represent <strong>CpG-rich genomic regions</strong> (including CpG islands, CpG island shores, enhancers, and other gene-regulatory elements), which are particularly relevant for epigenetic regulation. Similar to exome-sequencing for mutation discovery, the RRBS protocol enriches for some of the most interesting target regions and thereby achieves a reduction in sequencing cost of a factor of 10-20 compared to whole genome bisulfite sequencing.</div>
</div>
<table width="1039">
<tbody>
<tr></tr>
<tr>
<td width="281"> </td>
<td width="379">
<h4>Premium RRBS Kit</h4>
</td>
<td width="379">
<h5>Illumina® RRBS protocol</h5>
</td>
</tr>
<tr>
<td><strong>DNA input</strong></td>
<td>100 ng</td>
<td>2-5 µg</td>
</tr>
<tr>
<td><strong>Multiplexing</strong></td>
<td>Pooling of 6 samples (one HiSeq lane)</td>
<td>no guidelines</td>
</tr>
<tr>
<td><strong>Bisulfite conversion</strong></td>
<td>One bisulfite reaction per pool (6 samples)</td>
<td>One bisulfite reaction per sample</td>
</tr>
<tr>
<td><strong># purification steps</strong></td>
<td>2</td>
<td>7</td>
</tr>
<tr>
<td><strong>Key features</strong></td>
<td width="379">low input - cost-effective - optimized for high-throughput - complete kit</td>
<td width="379">high input - not optimized for high-throughput - reagents from different providers</td>
</tr>
</tbody>
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<p></p>
<h1 class="advertising-feature"><strong>Premium RRBS technology: cost-effective DNA methylation mapping with superior coverage</strong></h1>
<ul class="citation dates">
<li style="text-align: left;" class="vcard last-author"><a href="#auth-5" class="name"><span class="fn">Christoph Bock</span></a><sup><a href="#a2">2</a></sup><sup href="#affil-auth">, </sup></li>
<li style="text-align: left;" class="vcard"><a href="#auth-4" class="name"><span class="fn">Sharon Squazzo</span></a><sup><a href="#a3">3</a></sup><sup>, </sup></li>
<li style="text-align: left;" class="vcard"><a href="#auth-3" class="name"><span class="fn">Miklos Laczik</span></a><sup><a href="#a1">1</a></sup><sup>, </sup></li>
<li style="text-align: left;" class="vcard"><a href="#auth-2" class="name"><span class="fn">Paul Datlinger</span></a><sup><a href="#a2">2</a></sup><sup>, </sup></li>
<li style="text-align: left;" class="vcard c1"><a href="#auth-1" class="name"><span class="fn">Anne-Clémence Veillard</span></a><sup><a href="#a1">1</a></sup><sup>, </sup></li>
</ul>
<p>Nature Methods 13 (2016)</p>
<p>Published online <time datetime="2016-01-28">28 January 2016 </time></p>
<p></p>
<p><a href="http://www.nature.com/nmeth/journal/v13/n2/full/nmeth.f.391.html" class="alert button"><span style="font-weight: 400;">CHECK OUT THE NATURE METHODS PAPER</span></a></p>
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<div class="extra-spaced">
<p>メチル化アレイとWGBS技術の長所を有するReduced representation bisulfite sequencing (RRBS) 技術は、あらゆる脊椎動物種で柔軟に使用できるため、大規模コホートの研究においても正確で費用対効果の高いアッセイを行えます。制限MspI酵素（CCGGターゲットサイト）を使用してゲノムを切断し、次にサイズを選択することで、濃縮されたDNAはプロモーターやCpGアイランドを含む関連するターゲットCpGリッチ領域を表します。またRRBS技術はゲノムの一部分を濃縮してメチル化解析を行うことによってシーケンス量を減らす手法でカバレッジが非常に高く、CpGメチル化解析に適しています。（ヒトサンプルで最大700万CpGが検出）。</p>
<h2>Genome-scale DNA methylation analysis</h2>
<ul class="square">
<li>Robust and cost-effective solution with reliable results</li>
<li>High coverage - up to 7 million CpGs detected in human samples</li>
<li>Single nucleotide resolution</li>
<li>Detection of methylation patterns in CpG rich regions including promoters and CpG islands</li>
<li>Suitable for any vertebrate species</li>
</ul>
<h2>Complete end-to-end service</h2>
<ul class="square">
<li>From DNA QC to standard bioinformatics </li>
<li>Leveraging our <a href="https://www.diagenode.com/en/p/premium-rrbs-kit-V2-x24">Premium RRBS kit V2 </a>widely adopted</li>
<li>As little as 25 ng for any vertebrate species</li>
<li>Differentially methylated site analysis </li>
</ul>
</div>
<div class="extra-spaced">
<h2></h2>
</div>
<p><span><i class="fa fa-arrow-circle-right"></i> </span><a href="https://www.diagenode.com/en/categories/dna-methylation-profiling-services">See our other DNA Methylation Profiling Services</a></p>
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<h4>RRBS Service includes:</h4>
<table style="width: 905px;">
<tbody>
<tr>
<td style="width: 264px;"><strong>QC of the genomic DNA</strong></td>
<td style="width: 636px;">
<ul style="list-style-type: circle;">
<li>Measurement of DNA concentration </li>
<li>Assessment of DNA quality</li>
</ul>
</td>
</tr>
<tr>
<td style="width: 264px;"><strong>Preparation of RRBS libraries</strong></td>
<td style="width: 636px;">
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<li>MspI digestion</li>
<li>Library preparation (ends preparation, adaptor ligation)</li>
<li>Size selection</li>
<li>Sample pooling</li>
<li>Bisulfite conversion</li>
<li>Library amplification and clean-up</li>
<li>QC of the RRBS library pool (DNA concentration, analysis of the pool profile)</li>
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<tr>
<td style="width: 264px;"><strong>Deep sequencing</strong></td>
<td style="width: 636px;">
<ul style="list-style-type: circle;">
<li>Samples are sequenced on an Illumina platform, paired-end reads, read length 50 bp (PE50)</li>
<li>40 million raw reads (on average) per sample when pooling 10 samples/lane</li>
<li>7 million CpGs (on average) for human samples</li>
<li>7-11x CpG coverage (on average) for human samples</li>
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<h4><strong>Analysis</strong></h4>
</th>
<th style="width: 624px;">
<h4><strong>Features</strong></h4>
</th>
</tr>
</thead>
<tbody>
<tr>
<td style="width: 262px;"><strong>Standard</strong></td>
<td style="width: 624px;">
<ul>
<li>FASTQ raw data</li>
<li>FASTQC quality control insights</li>
<li>Alignment of bisulfite sequencing data against reference genome</li>
<li>Methylation calling and extraction</li>
<li>Summary statistics</li>
</ul>
</td>
</tr>
<tr>
<td style="width: 262px;"><strong>Differential methylation analysis<br /></strong></td>
<td style="width: 624px;">
<ul>
<li>Methylation level analysis</li>
<li>Differentially Methylated CpGs (DMCs) analysis</li>
<li>Differentially Methylated Regions (DMRs) analysis</li>
<li>Annotation of DMCs and DMRs for genomic regions (exons, introns, …)</li>
<li>Clustering analysis</li>
</ul>
</td>
</tr>
<tr>
<td style="width: 262px;">
<p><strong>Gene ontology terms analysis</strong></p>
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<td style="width: 624px;">
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<li>Enrichment analysis on gene associated with DMCs and DMRs</li>
<li>Get functional insights</li>
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<tr>
<td style="width: 262px;">
<p><strong>Pathway analysis</strong></p>
</td>
<td style="width: 624px;">
<ul>
<li>Identification of biological pathways in which genes associated with DMCs and DMRs may be over-represented (or under-represented)</li>
<li>Get mechanistic insights</li>
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<div style="text-align: justify;" class="large-12 columns">Bisulfite modification of DNA is the most commonly used, "<strong>gold standard</strong>" method for DNA methylation studies providing <strong>single nucleotide resolution</strong>. T<span style="font-weight: 400;">his technology is based on the chemical conversion of unmethylated cytosine to uracil. Methylated cytosines are protected from this conversion allowing to determine DNA methylation at the singe nucleotide level.</span></div>
<div style="text-align: justify;" class="large-12 columns"></div>
<div style="text-align: justify;" class="large-12 columns">Various analyses can be performed on the altered sequence to retrieve this information: bisulfite sequencing, pyrosequencing, methylation-specific PCR, high resolution melting curve analysis, microarray-based approaches, and next-generation sequencing.
<h3>How it works</h3>
Treatment of DNA with bisulfite converts cytosine residues to uracil, but leaves 5-methylcytosine residues unaffected (see Figure 1).
<p class="text-center"><img src="https://www.diagenode.com/img/applications/bisulfite.png" /><br />Figure 1: Overview of bisulfite conversion of DNA</p>
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<h2>Complete solutions for DNA methylation studies</h2>
<p>Whether you are experienced or new to the field of DNA methylation, Diagenode has everything you need to make your assay as easy and convenient as possible while ensuring consistent data between samples and experiments. Diagenode offers sonication instruments, reagent kits, high quality antibodies, and high-throughput automation capability to address all of your specific DNA methylation analysis requirements.</p>
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<p>DNA methylation was the first discovered epigenetic mark and is the most widely studied topic in epigenetics. <em>In vivo</em>, DNA is methylated following DNA replication and is involved in a number of biological processes including the regulation of imprinted genes, X chromosome inactivation. and tumor suppressor gene silencing in cancer cells. Methylation often occurs in cytosine-guanine rich regions of DNA (CpG islands), which are commonly upstream of promoter regions.</p>
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<div class="small-12 medium-12 large-12 columns"><br /><br />
<ul class="accordion" data-accordion="">
<li class="accordion-navigation"><a href="#dnamethyl"><i class="fa fa-caret-right"></i> Learn more</a>
<div id="dnamethyl" class="content">5-methylcytosine (5-mC) has been known for a long time as the only modification of DNA for epigenetic regulation. In 2009, however, Kriaucionis discovered a second methylated cytosine, 5-hydroxymethylcytosine (5-hmC). The so-called 6th base, is generated by enzymatic conversion of 5-methylcytosine (5-mC) into 5-hydroxymethylcytosine by the TET family of oxygenases. Early reports suggested that 5-hmC may represent an intermediate of active demethylation in a new pathway which demethylates DNA, converting 5-mC to cytosine. Recent evidence fuel this hypothesis suggesting that further oxidation of the hydroxymethyl group leads to a formyl or carboxyl group followed by either deformylation or decarboxylation. The formyl and carboxyl groups of 5-formylcytosine (5-fC) and 5-carboxylcytosine (5-caC) could be enzymatically removed without excision of the base.
<p class="text-center"><img src="https://www.diagenode.com/img/categories/kits_dna/dna_methylation_variants.jpg" /></p>
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</li>
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<br />
<h2>Main DNA methylation technologies</h2>
<p style="text-align: justify;">Overview of the <span style="font-weight: 400;">three main approaches for studying DNA methylation.</span></p>
<div class="row">
<ol>
<li style="font-weight: 400;"><span style="font-weight: 400;">Chemical modification with bisulfite – Bisulfite conversion</span></li>
<li style="font-weight: 400;"><span style="font-weight: 400;">Enrichment of methylated DNA (including MeDIP and MBD)</span></li>
<li style="font-weight: 400;"><span style="font-weight: 400;">Treatment with methylation-sensitive or dependent restriction enzymes</span></li>
</ol>
<p><span style="font-weight: 400;"> </span></p>
<div class="row">
<table>
<thead>
<tr>
<th></th>
<th>Description</th>
<th width="350">Features</th>
</tr>
</thead>
<tbody>
<tr>
<td><strong>Bisulfite conversion</strong></td>
<td><span style="font-weight: 400;">Chemical conversion of unmethylated cytosine to uracil. Methylated cytosines are protected from this conversion allowing to determine DNA methylation at single nucleotide resolution.</span></td>
<td>
<ul style="list-style-type: circle;">
<li style="font-weight: 400;"><span style="font-weight: 400;">Single nucleotide resolution</span></li>
<li style="font-weight: 400;"><span style="font-weight: 400;">Quantitative analysis - methylation rate (%)</span></li>
<li style="font-weight: 400;"><span style="font-weight: 400;">Gold standard and well studied</span></li>
<li style="font-weight: 400;"><span style="font-weight: 400;">Compatible with automation</span></li>
</ul>
</td>
</tr>
<tr>
<td><b>Methylated DNA enrichment</b></td>
<td><span style="font-weight: 400;">(Hydroxy-)Methylated DNA is enriched by using specific antibodies (hMeDIP or MeDIP) or proteins (MBD) that specifically bind methylated CpG sites in fragmented genomic DNA.</span></td>
<td>
<ul style="list-style-type: circle;">
<li style="font-weight: 400;"><span style="font-weight: 400;">Resolution depends on the fragment size of the enriched methylated DNA (300 bp)</span></li>
<li style="font-weight: 400;"><span style="font-weight: 400;">Qualitative analysis</span></li>
<li style="font-weight: 400;"><span style="font-weight: 400;">Compatible with automation</span></li>
</ul>
</td>
</tr>
<tr>
<td><strong>Restriction enzyme-based digestion</strong></td>
<td><span style="font-weight: 400;">Use of (hydroxy)methylation-sensitive or (hydroxy)methylation-dependent restriction enzymes for DNA methylation analysis at specific sites.</span></td>
<td>
<ul style="list-style-type: circle;">
<li style="font-weight: 400;"><span style="font-weight: 400;">Determination of methylation status is limited by the enzyme recognition site</span></li>
<li style="font-weight: 400;"><span style="font-weight: 400;">Easy to use</span></li>
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<p>Sodium bisulfite conversion of genomic DNA is the most commonly used method for DNA methylation studies providing <strong>single nucleotide resolution</strong>. It enables <span>to differentiate and detect unmethylated versus methylated cytosines. This procedure can then be followed either by <strong>PCR amplification</strong> or <strong>next generation sequencing</strong> to reveal the methylation status of every cytosine in gene specific amplification or whole genome amplification.</span></p>
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<h2>How it works</h2>
<p style="text-align: left;">Treatment of DNA with sodium bisulfite converts unmethylated cytosine to uracil, while methylated cytosines remain unchanged. <span>The DNA is then amplified by PCR where the uracils are converted to thymines. </span></p>
<p style="text-align: center;"><span></span></p>
<p><img src="https://www.diagenode.com/img/categories/bisulfite-conversion/bisulfite-conversion-acgautac.png" style="display: block; margin-left: auto; margin-right: auto;" /></p>
<h2>Advantages</h2>
<ul class="nobullet" style="font-size: 19px;">
<li><i class="fa fa-arrow-circle-right"></i><strong> </strong><strong>Single nucleotide</strong> resolution</li>
<li><i class="fa fa-arrow-circle-right"></i><strong> Gene-specific </strong>and <strong>genome-wide</strong><span> analyses</span></li>
<li><i class="fa fa-arrow-circle-right"></i><strong> NGS</strong><span> </span>compatible</li>
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<h2>Downstream analysis techniques</h2>
<ul class="square">
<li>Reduced Representation Bisulfite Sequencing (RRBS) with our <a href="https://www.diagenode.com/en/p/premium-rrbs-kit-V2-x24">Premium RRBS Kit V2</a></li>
<li>Bisulfite conversion with our <a href="https://www.diagenode.com/en/p/premium-bisulfite-kit-50-rxns">Premium Bisulfite Kit</a> followed by qPCR, Sanger, Pyrosequencing</li>
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<p><span>Positive and negative spike-in controls are included for the monitoring of bisulfite conversion efficiency. </span></p>
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<p><span>Size selection has been optimized to keep small fragments of interest and to remove adaptor dimers, resulting in a better coverag. </span></p>
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<p><span>The pooling protocol includes a quantification of the samples and a pooling application, available on our website, to help you to choose the groups, depending on the DNA amount and adaptor barcode of each sample. </span></p>
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<p><span>The bisulfite conversion protocol has been improved to decrease DNA degradation while keeping a highly efficient conversion of unmethylated cytosine. </span></p>
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<h6 style="height:60px">Premium RRBS kit (24 rxns) - replaced by Premiu...</h6>
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<a href="/en/p/premium-rrbs-kit-x96-96-rxns"><img src="/img/product/kits/methyl-kit-icon.png" alt="Methylation kit icon" class="th"/></a> </div>
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<h6 style="height:60px">Premium RRBS kit (96 rxns) - replaced by Premiu...</h6>
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<h3>Get a quote</h3><p class="lead">You are about to request a quote for <strong>our epigenomics services</strong>. Fill out the form below and we will be in touch with you very soon.</p><p><small>All <span style="font-size:16px;color:red;">*</span> fields are mandatory</small></p>
</div>
</div>
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<h2>Service Information</h2>
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<div class="small-12 large-12 columns">
<h4>Which services are you interested in?</h4>
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<div class="small-12 large-12 columns">
<input type="hidden" name="data[Quote][epigenomics_service]" value="" id="QuoteEpigenomicsService"/>
<div class="checkbox"><input type="checkbox" name="data[Quote][epigenomics_service][]" value="ChIP-seq" id="QuoteEpigenomicsServiceChIPSeq" /><label for="QuoteEpigenomicsServiceChIPSeq">ChIP-seq</label></div>
<div class="checkbox"><input type="checkbox" name="data[Quote][epigenomics_service][]" value="ATAC-seq" id="QuoteEpigenomicsServiceATACSeq" /><label for="QuoteEpigenomicsServiceATACSeq">ATAC-seq</label></div>
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<div class="checkbox"><input type="checkbox" name="data[Quote][epigenomics_service][]" value="WGBS" id="QuoteEpigenomicsServiceWGBS" /><label for="QuoteEpigenomicsServiceWGBS">WGBS</label></div>
<div class="checkbox"><input type="checkbox" name="data[Quote][epigenomics_service][]" value="MeDIP-seq" id="QuoteEpigenomicsServiceMeDIPSeq" /><label for="QuoteEpigenomicsServiceMeDIPSeq">MeDIP-seq</label></div>
<div class="checkbox"><input type="checkbox" name="data[Quote][epigenomics_service][]" value="Targeted DNA methylation analysis" id="QuoteEpigenomicsServiceTargetedDNAMethylationAnalysis" /><label for="QuoteEpigenomicsServiceTargetedDNAMethylationAnalysis">Targeted DNA methylation analysis</label></div>
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<div class="checkbox"><input type="checkbox" name="data[Quote][epigenomics_service][]" value="Infinium Mouse Methylation Array" id="QuoteEpigenomicsServiceInfiniumMouseMethylationArray" /><label for="QuoteEpigenomicsServiceInfiniumMouseMethylationArray">Infinium Mouse Methylation Array</label></div>
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<div class="checkbox"><input type="checkbox" name="data[Quote][epigenomics_service][]" value="Data mining" id="QuoteEpigenomicsServiceDataMining" /><label for="QuoteEpigenomicsServiceDataMining">Data mining</label></div>
<div class="checkbox"><input type="checkbox" name="data[Quote][epigenomics_service][]" value="Human Methylome" id="QuoteEpigenomicsServiceHumanMethylome" /><label for="QuoteEpigenomicsServiceHumanMethylome">Human Methylome</label></div>
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<div class="row collapse">
<div class="small-12 medium-12 large-3 columns">
<span class="prefix">Sample species</span>
</div>
<div class="small-12 medium-12 large-9 columns">
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</div>
<div class="row collapse">
<div class="small-12 medium-12 large-6 columns">
<span class="prefix">Total number of samples (including replicates)</span>
</div>
<div class="small-12 medium-12 large-6 columns">
<input name="data[Quote][number_samples]" maxlength="255" type="text" id="QuoteNumberSamples"/> </div>
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<div class="row collapse">
<h2>Contact Information</h2>
<div class="small-3 large-2 columns">
<span class="prefix">First name <sup style="font-size:16px;color:red;">*</sup></span>
</div>
<div class="small-9 large-10 columns">
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<span class="prefix">Company <sup style="font-size:16px;color:red;">*</sup></span>
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<span class="prefix">Phone number</span>
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<div class="small-9 large-10 columns">
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<div class="small-3 large-2 columns">
<span class="prefix">City</span>
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<option value="AF">Afghanistan</option>
<option value="AX">Åland Islands</option>
<option value="AL">Albania</option>
<option value="DZ">Algeria</option>
<option value="AS">American Samoa</option>
<option value="AD">Andorra</option>
<option value="AO">Angola</option>
<option value="AI">Anguilla</option>
<option value="AQ">Antarctica</option>
<option value="AG">Antigua and Barbuda</option>
<option value="AR">Argentina</option>
<option value="AM">Armenia</option>
<option value="AW">Aruba</option>
<option value="AU">Australia</option>
<option value="AT">Austria</option>
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<option value="BW">Botswana</option>
<option value="BV">Bouvet Island</option>
<option value="BR">Brazil</option>
<option value="IO">British Indian Ocean Territory</option>
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<option value="BG">Bulgaria</option>
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<option value="BI">Burundi</option>
<option value="KH">Cambodia</option>
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<option value="CV">Cape Verde</option>
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<option value="CF">Central African Republic</option>
<option value="TD">Chad</option>
<option value="CL">Chile</option>
<option value="CN">China</option>
<option value="CX">Christmas Island</option>
<option value="CC">Cocos (Keeling) Islands</option>
<option value="CO">Colombia</option>
<option value="KM">Comoros</option>
<option value="CG">Congo</option>
<option value="CD">Congo, The Democratic Republic of the</option>
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<option value="CR">Costa Rica</option>
<option value="CI">Côte d'Ivoire</option>
<option value="HR">Croatia</option>
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<option value="CW">Curaçao</option>
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<option value="DJ">Djibouti</option>
<option value="DM">Dominica</option>
<option value="DO">Dominican Republic</option>
<option value="EC">Ecuador</option>
<option value="EG">Egypt</option>
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<option value="GQ">Equatorial Guinea</option>
<option value="ER">Eritrea</option>
<option value="EE">Estonia</option>
<option value="ET">Ethiopia</option>
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<option value="GM">Gambia</option>
<option value="GE">Georgia</option>
<option value="DE">Germany</option>
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<option value="GD">Grenada</option>
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<h6 style="height:60px">RRBS Service (Reduced Representation Bisulfite ...</h6>
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<p>メチル化アレイとWGBS技術の長所を有するReduced representation bisulfite sequencing (RRBS) 技術は、あらゆる脊椎動物種で柔軟に使用できるため、大規模コホートの研究においても正確で費用対効果の高いアッセイを行えます。制限MspI酵素（CCGGターゲットサイト）を使用してゲノムを切断し、次にサイズを選択することで、濃縮されたDNAはプロモーターやCpGアイランドを含む関連するターゲットCpGリッチ領域を表します。またRRBS技術はゲノムの一部分を濃縮してメチル化解析を行うことによってシーケンス量を減らす手法でカバレッジが非常に高く、CpGメチル化解析に適しています。（ヒトサンプルで最大700万CpGが検出）。</p>
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<li>Single nucleotide resolution</li>
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<h2>Complete end-to-end service</h2>
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<li>Leveraging our <a href="https://www.diagenode.com/en/p/premium-rrbs-kit-V2-x24">Premium RRBS kit V2 </a>widely adopted</li>
<li>As little as 25 ng for any vertebrate species</li>
<li>Differentially methylated site analysis </li>
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<h2></h2>
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<p><span><i class="fa fa-arrow-circle-right"></i> </span><a href="https://www.diagenode.com/en/categories/dna-methylation-profiling-services">See our other DNA Methylation Profiling Services</a></p>
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<li>7-11x CpG coverage (on average) for human samples</li>
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<h4><strong>Analysis</strong></h4>
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<h4><strong>Features</strong></h4>
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<li>Get functional insights</li>
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