Diagenode

Auto True MicroChIP kit

Catalog Number
Format
Price
C01010140
16 rxns
$590.00

The Auto True MicroChIP kit in combination with the MicroPlex Library Preparation™ kit allows for performing ChIP-seq on as few as 10,000 cells. This microChIP-seq assay has been validated with the new generation IP-Star® Compact Automated Workstation. Furthermore Diagenode's high quality ChIP-seq grade antibodies have been used.

Video article

Automating ChIP-seq Experiments to Generate Epigenetic Profiles on 10,000 HeLa Cells

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  • Characteristics
    • Revolutionary: Only 10.000 cells needed for complete ChIP-seq procedure
    • Reliable and accurate sequencing library preparation from just picogram inputs
    • Validated on studies for histone marks and with the IP-Star® Compact Automated Platform

    ChIP on 10,000 cells

    High efficiency ChIP on 10,000 cells
    ChIP efficiency on 10 000 cells. ChIP was performed on human Hela cells using the Diagenode antibodies H3K4me3 (Cat. No. pAb-003-050), H3K27ac (pAb-174-050), H3K9me3 (pAb-056-050) and H3K27me3 (pAb-069-050). Sheared chromatin from 10 000 cells and 0.1 µg (H3K27ac), 0.25 µg (H3K4me3 and H3K27me3) or 0.5 µg (H3K9me3) of the antibody were used per IP. Corresponding amount of IgG was used as control. Quantitative PCR was performed with primers for corresponding positive and negative loci. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

    ChIP amplification

    Match between True MicroChIP peaks and the reference dataset

    Reliable detection of enrichments in ChIP-seq
    A: ChIP has been peformed with H3K4me3 antibody, amplification of 17 pg of DNA ChIP'd from 10.000 cells and amplification of 35 pg of DNA ChIP'd from 100.000 cells (control experiment). The IP'd DNA was amplified and transformed into a sequencing-ready preparation for the Illumina plateform with the MicroPlex Library Preparation kit. The library was then analysed on an Illumina® Genome Analyzer. Cluster generation and sequencing were performed according to the manufacturer's instructions.
    B: We observed a perfect match between the top 40% of True MicroChIP peaks and the reference dataset. Based on the NIH Encode project criterion, ChIP-seq results are considered reproducible between an original and reproduced dataset if the top 40% of peaks have at least an 80% overlap ratio with the compared dataset.

    Cell lysis

    Hela cells were fixed with 1% formaldehyde (for 10 minutes at RT)
    Cell lysis was performed using the Lysis Buffer tL1 of the Diagenode True MicroChIP kit. Samples corresponding to 10 000 cells are sheared during 5 rounds of 5 cycles of 30 seconds “ON” / 30 seconds “OFF” with the Bioruptor® Plus combined with the Bioruptor® Water cooler (Cat No. BioAcc-cool) at HIGH power setting (position H). For optimal results, samples are vortexed before and after performing 5 sonication cycles, followed by a short centrifugation at 4°C. 10 μl of DNA (equivalent to 60 000 cells) are analysed on a 1.5% agarose gel.

  •  Documents
    Auto TrueMicroChIP kit MANUAL
    Conventional ChIP protocols require high numbers of cells (hundreds of thousands cells at l...
    Download
    Chromatin Brochure BROCHURE
    Whether you are experienced or new to the field of chromatin immunoprecipitation, Diagenode has e...
    Download
  •  Publications

    How to properly cite this product in your work

    Diagenode strongly recommends using this: Auto True MicroChIP kit (Diagenode Cat# C01010140). Click here to copy to clipboard.

    Using our products in your publication? Let us know!

    EZH2 is overexpressed in transitional preplasmablasts and is involved in human plasma cell differentiation.
    Herviou L, Jourdan M, Martinez AM, Cavalli G, Moreaux J
    Plasma cells (PCs) play a major role in the defense of the host organism against pathogens. We have shown that PC generation can be modeled using multi-step culture systems that reproduce the sequential cell differentiation occurring in vivo. Using this unique model, we investigated the role of EZH2 during PC differ...

    An Integrated and Semiautomated Microscaled Approach to Profile Cis-Regulatory Elements by Histone Modification ChIP-Seq for Large-Scale Epigenetic Studies.
    Youhanna Jankeel D, Cayford J, Schmiedel BJ, Vijayanand P, Seumois G.
    Chromatin immunoprecipitation followed by sequencing (ChIP-Seq) is the preferred approach to map histone modifications and identify cis-regulatory DNA elements throughout the genome. Multiple methods have been described to increase the efficiency of library preparation and to reduce hands-on time as well as costs. T...

    Reciprocal signalling by Notch-Collagen V-CALCR retains muscle stem cells in their niche.
    Baghdadi MB, Castel D, Machado L, Fukada SI, Birk DE, Relaix F, Tajbakhsh S, Mourikis P
    The cell microenvironment, which is critical for stem cell maintenance, contains both cellular and non-cellular components, including secreted growth factors and the extracellular matrix. Although Notch and other signalling pathways have previously been reported to regulate quiescence of stem cells, the co...

    A new metabolic gene signature in prostate cancer regulated by JMJD3 and EZH2.
    Daures M, Idrissou M, Judes G, Rifaï K, Penault-Llorca F, Bignon YJ, Guy L, Bernard-Gallon D
    Histone methylation is essential for gene expression control. Trimethylated lysine 27 of histone 3 (H3K27me3) is controlled by the balance between the activities of JMJD3 demethylase and EZH2 methyltransferase. This epigenetic mark has been shown to be deregulated in prostate cancer, and evidence shows H3K27me3 enri...

    Global analysis of H3K27me3 as an epigenetic marker in prostate cancer progression
    Ngollo M. et al.
    BACKGROUND: H3K27me3 histone marks shape the inhibition of gene transcription. In prostate cancer, the deregulation of H3K27me3 marks might play a role in prostate tumor progression. METHODS: We investigated genome-wide H3K27me3 histone methylation profile using chromatin immunoprecipitation (ChIP) and 2X400K p...

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