CpG MethylQuest DNA Isolation Kit

Methylation of cytosines located 5′ to guanosine is known to have a profound effect on the expression of many eukaryotic genes. In normal cells methylation occurs predominantly in CG-poor regions, while CG-rich areas, called CpG-islands remain unmethylated. The exceptions are the extensive methylation of CpG islands associated with transcriptional inactivation of regulatory regions of imprinted genes and genes on the inactive X-chromosome of females. Aberrant methylation of normally unmethylated CpG islands has been documented as a relatively frequent event in immortalized and transformed cells and has been associated with transcriptional inactivation of defined tumor suppressor genes in human cancers.

To evalute the methylation status of either a specific locus or an entire genome, the isolation and enrichment of methylated DNA can be a useful first step. To enable this enrichement, the CpG MethylQuest DNA Isolation Kit utilizes a high-affinity GST-MBD protein pre-bound to a magnetic bead.

This purified recombinant protein contains the methyl binding domain (MBD) of the mouse MBD2b protein fused to a glutathione-S-transferase protein (GST) from S. japonicum separated by a linker containing a thrombin cleavage site. The MBD from the MBD2b protein has the highest affinity among the known methyl CpG binding proteins for Me-CpG sites and the lowest cross reactivity with unmethylated CpG sequences (Fraga M.F., et al. (2003). Nuc. Acids Res, 31, 1765–1774). These properties allow for very specific enrichment of methylated regions.

In addition to its high affinity for methylated DNA, the CpG MethylQuest protein binds to methylated CpG regardless of sequence context. This is in contrast to isolation of methylated DNA using proteins such as MeCP2, which requires a run of A-Ts near a CpG site. This combination of sequence independent, high affinity binding to methylated CpG sites and extremely low affinity for non-methylated sites results in a greater number of methylated CpG sites being recognized allowing for comprehensive analysis of methylation patterns across the genome.

The CpG MethylQuest DNA Isolation Kit allows for simple, effective and rapid enrichment of methylated DNA from genomic samples. Featuring a high affinity recombinant methyl-binding domain (MBD) protein pre-bound to magnetic beads, a rapid protocol, and a simple elution procedure that yields methylated DNA ready for downstream analysis, this high affinity kit enriches for methylated DNA from as little as 1 ng of DNA!

CpG MethylQuest Advantages

• Specific enrichment of methylated DNA fragments
• No detectable binding of unmethylated or hemimethylated regions
• Simple and fast 2-hour magnetic bead-based protocol
• GST-MBD2b capture protein pre-bound to magnetic beads for consistent results
• Reliable performance from 1 ng to 1 μg of DNA
• Elute ready-to-use DNA & avoid additional cleanup steps that reduce yields

Research Category
Epigenetics & Nuclear Function

CpG MethylQuest Binding Buffer

CpG MethylQuest Magnetic Beads with pre-bound recombinant MBD protein

CpG MethylQuest Wash Buffer 1

CpG MethylQuest Wash Buffer 2

TE Buffer

Control HeLa DNA 110 ng, MseI cut, 2ng/µl

Positive Control Primers 10 µM each

Negative Control Primers 20 µM each

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

CPG METHYLQUEST is a registered trademark of Merck KGaA, Darmstadt, Germany

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.