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NEB/EpiMark? Methylated DNA Enrichment Kit/25 reactions/E2600S
  • NEB/EpiMark? Methylated DNA Enrichment Kit/25 reactions/E2600S

NEB/EpiMark? Methylated DNA Enrichment Kit/25 reactions/E2600S

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貨號(hào): E2600S
品牌: NEB
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    • The EpiMark® Methylated DNA Enrichment Kit will selectively bind and enrich double-stranded methyl-CpG DNA from fragmented genomic DNA with as little as 5 ng of input DNA.Methylated DNA is isolated from fragmented genomic DNA (5 ng–25 μg) by binding to the methyl-CpG binding domain of human MBD2 protein fused to the Fc tail of human IgG1 (MBD2-Fc), which is coupled to paramagnetic hydrophilic protein A beads (MBC2-Fc/Protein A Magnetic Bead). Two Fc domains can be bound to one site on protein A with high affinity (Kd=10-7). As the Fc fragment is a dimer, four MBD2 domains are exposed to the solvent per molecule of protein A, increasing the relative equilibrium constant 100-fold. This stable complex will selectively bind double-stranded methylated CpG containing DNA. After simple wash steps followed by magnetic capture, the enriched DNA sample is easily eluted in a small volume of nuclease-free water by incubation at 65°C. The sample is immediately ready for downstream analysis by a variety of methods including:
      • Endpoint and real-time PCR assays
      • Bisulfite conversion followed by DNA amplification
      • Cloning and sequencing
      • Direct sequencing
      • Library preparation for high-throughput sequencing
      • Labeling for DNA microarray analysis
      • Methylation-sensitive restriction enzyme-based assays

      MBD-Fc and MCIp were originally developed by Michael Rehli at the University of Regensburg to improve the sensitivity and specificity of conventional CpG binding techniques.Kit Components:Each kit contains sufficient reagents for the enrichment of methylated DNA from up to 100 μg of fragmented input DNA. If starting with 5 ng to 10 μg of input DNA per experiment, the kit provides sufficient reagents for 25 reactions. Store at 4°C. For long-term storage > 6 months, MBD2-Fc protein should be stored at -20°C.

      Figure 1. Enrichment workflow
      Figure 2: Endpoint PCR of a methylated locus (LINE) that has been purified using the EpiMark methylated DNA Enrichment Kit
      Analyzed fractions are described above gel.
      This product is related to the following categories:
      Methylome Analysis,
      Epigenetic Analysis,
      Next Generation Sequencing Library Preparation
      This product can be used in the following applications:
      DNA Enrichment
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