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MethylAffinity™ Methylated DNA Enrichment Kit

MethylAffinity™ Methylated DNA enrichment kit provides a GCM™ bead-based method for quick enrichment of methylated DNA fragments from whole genome. The enriched sample improves the performance of downstream analysis, such as RT-PCR, microarray and sequencing for methylation status and location studies.

The genetically engineered GCM recombinant protein is designed based on the functional domain of methyl binding domain (MBD) proteins. It binds specifically to double-stranded DNA containing methylated CpG dinucleotides and has much higher affinity and specificity than any wild-type MBDs.

Protocol overview

Advantages

Fewer steps and less time

  • Whole process takes < 2 hours
  • Immobilized GCM magenta beads are provided. No need to perform beads-protein coupling reaction.
  • GCM captures double-stranded methylated DNA, which simplifies genomic sample preparation and downstream analysis.
  • Rapid elution with salt.

High affinity

  • Higher affinity than other commercially available MBD proteins.
  • Be able to isolate DNA fragments which contain only a few copies of methylated CpG dinucleotides.
  • Be able to enrich methylated DNA from nanograms of genomic DNA sample.

Gradient elution option

  • For precise study of methylation status, salt gradient elution can be used to fractionate DNA fragments based on methylated-CpG density.

Convenience

  • Vivid magenta color makes the beads easy to spot, thus greatly reduces the risk of incidental beads loss.
  • Restriction enzymes are provided in the kit for genomic DNA fragmentation.
 
Figure 1. PCR results of enrichment controls 100 ng of mouse genomic DNA (sonicated) were added to 4 µl of GCM-beads. After 1 hour binding reaction at RT, the beads were washed several times. Then the methylated DNA were eluted from the beads. The wash and elution supernatants (5% of each) were analyzed by PCR using primer sets specific for mouse β-actin promoter (unmethylated), GAPDH promoter (unmethylated), and IAP repeats (methylated). The PCR products were checked by agarose gel electrophoresis.   Figure 2. Easy to spot GCM beads Comparison of GCM magenta beads (right) with conventional Sepharose beads (left). The magenta color of the beads helps reduce accidental beads loss during the handling.

Buy Catalog# Product Description Price
FF003 MethylAffinity™ Methylated DNA Enrichment Kit
(30 reactions)
GCM-beads based method for quick enrichment of double-stranded DNA fragments containing methylated CpG dinucleotides from whole genome. inquire

 

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Background of DNA Methylation

DNA methylation in mammals is involved in many cellular processes and is responsible for gene silencing by recruitment of transcriptional repression complexes. It plays a critical role in development. However, massive methylation of the promoter regions has been frequently observed in cancer, which can cause transcription repression of tumor suppressor and DNA repair genes.