CpG MethylQuest DNA Isolation Kit Chemische Eigenschaften,Einsatz,Produktion Methoden
Verwenden
Research Category
Epigenetics & Nuclear Function
Allgemeine Beschreibung
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.
CpG MethylQuest DNA Isolation Kit Upstream-Materialien And Downstream Produkte
Upstream-Materialien
Downstream Produkte
