The CpGenome™ Human Methylated DNA Standard is purified from HCT116 DKO cells that contain genetic knockouts of DNA methyltransferases, DNMT1 (-/-) and DNMT3b (-/-) , and have less than 5% methylated DNA. This Standard has been methylated enzymatically at all CpG dinucleotides by M. SssI methyltransferase 2. It is intended for use as a positive control in gene methylation studies, such as bisulfite conversion of DNA with the CpGenome™ Turbo Bisulfite Modification Kit (Cat. No. S7847). Bisulfite-modified DNA can be evaluated by methylation-specific PCR (MSP) technology using the CpG WIZ® amplification kits.
One vial containing 5 µg (20 µL) of CpGenome™ Human Methylated DNA Standard at a concentration of 250 ng/µL.
Methylation-specific PCR (MSP) was performed on the CpGenome™ Human Methylated DNA after bisulfite modification with the CpGenome™ Turbo Bisulfite kit (Cat. No. S7847). Three sets of primers from the CpG WIZ® BRCA1 amplification kit (Cat. No. S7830) were used in this assay: the U primer set, which anneals to unmethylated bisulfite-modified DNA; M primers, which anneal to methylated bisulfite-modified sequences; and W primers, which anneal to unmethylated or methylated DNA that has not undergone bisulfite modification. Only the M primers generated product for the methylated DNA standard.View CpGenome™ Human Methylated DNA Standard on Millipore’s website