Bioruptor® in Action Video
Applications | Methylbinding domain protein
Methylated DNA enrichment with MBD (methylbinding domain) protein
DNA is typically modified with the addition of a methyl group at the fifth position of cytosine following DNA replication. DNA methylation is involved in a number of biological processes including 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.
The MBD approach is based on the very high affinity of a H6-GST-MBD fusion protein for methylated DNA. This protein consists of the methyl binding domain (MBD) of human MeCP2, as a C-terminal fusion with Glutathione-S-transferase (GST) containing an N-terminal His6-tag. The H6-GST-MBD fusion protein can be used to specifically isolate DNA containing methylated CpGs.
Diagenode’s MethylCap® kit enables high enrichment of double-stranded DNA and a differential fractionation in function of the methylated CpG density. Fractionation reduces the complexity of your sample and makes therefore subsequent next generation sequencing easier.
Prior to the MethylCap assay, DNA is first extracted and sheared using the Bioruptor® Sonicator.
|Auto MethylCap kit x48|
|MethylCap kit x48|
|H6-GST-MBD fusion protein|