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    Methylated-DNA IP Kit


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    Cat # Name Size
    D5101 Methylated-DNA IP Kit 10 Rxns.

    Highlights

    • Robust enrichment & immunoprecipitation of DNA containing 5mC.
    • Includes a highly specific anti-5-methylcytosine monoclonal antibody for defined, reproducible results.
    • Eluted, ultra-pure DNA is ideal for use in subsequent molecular-based analyses (e.g., assembling genomic libraries and determining genome-wide methylation status).
    Description & Documents Specifications Components Citations

    Documents

    Product Description

    The Methylated-DNA IP Kit is designed for enrichment of 5mC-containing DNA from any pool of fragmented genomic DNA for use in genome-wide methylation analysis. It features a highly specific Anti-5-Methylcytosine Monoclonal Antibody for the immunoprecipitation of methylated DNA in only a few hours. This kit is capable of achieving over one hundred-fold enrichment of methylated DNA vs. non-methylated DNA. Recovered DNA is suitable for many downstream applications to analyze genome-wide DNA methylation including PCR, bisulfite treatment, whole-genome amplification, ultra-deep sequencing, and microarray. Control DNA and primers are included to monitor the success of the assay.

    Documents

    Technical Specifications

    Applicable For Immunoprecipitation of methylated DNA, PCR, sequencing, microarrays, etc.
    Elution Volume ≥ 15 µl
    Input 50 - 500 ng of DNA
    Processing Time 4 hours

    Resources

    Cat # Name Size
    D5101-3-20 MIP Buffer 20 ml
    D5101-4-1 DNA Denaturing Buffer 1 ml
    A3002-50 Anti-5-Methylcytosine Monoclonal Antibody (Clone 7D21) 50 µl
    D5101-2 Methylated/Non-methylated Control DNA & Primer Set 1 Set
    D3004-4-10 DNA Elution Buffer 10 ml

    The authors utilized the DNA Methylation IP Kit (MeDIP) from Zymo Research to compare DNA methylation results using either MeDIP-Seq or Infinium HumanMethylation450 BeadChIP array. They found the MeDIP-Seq resulted in detection of 15,709 differentially methylation regions, almost twice as many compared the array based method (8070), and concluded that MeDIP-Seq provided a more comprehensive picture of the methylation in human samples.

    Clark C, Palta P, Joyce CJ, Scott C, Grundberg E, Deloukas P, Palotie A, Coffey A. 2012. A Comparison of the Whole Genome Approach of MeDIP-Seq to the Targeted Approach of the Infinium HumanMethylation450 BeadChIP for Methylome Profiling. PLOS ONE 7:11 (e5

    Researchers used the Methylated DNA IP Kit from Zymo Research to immunoprecipitate methylated DNA from leaf tissue and study the DNA methylation profile in the maize genome. The authors found that specific families of retrotransposons could spread heterochromatin to neighboring genes and epigenetically suppress their expression.

    Eichten SR et al. (2012) Spreading of heterochromatin is limited to specific families of maize retrotransposons. PLoS Genet. 8(12):e1003127.

    The Methylated-DNA IP Kit was used for CpG immunoprecipitation of genomic DNA isolated from T cells that was sheared to ~200 bp using dsDNA Shearase Plus (E2018) to study the DNA methylation status of the Notch-1 promoter.

    Rauen T et al. (2012) AMP-responsive element modulator α (CREMα) contributes to decreased Notch-1 expression in T cells from patients with active systemic lupus erythematosus (SLE). J Biol Chem. 287(51):42525-32.

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