CpGenome™ Human Methylated & Non-methylated DNA Standards are purified from HCT116 DKO cells that contain genetic knockouts of DNA methyltransferases, DNMT1 (-/-) and DNMT3b (-/-) , and have less than 5% methylated DNA. The CpGenome™ Human Methylated DNA Standard has been methylated enzymatically at all CpG dinucleotides by M. SssI methyltransferase 2. These standards are intended for use as positive and negative controls 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.
Two vials, each containing 5 µg (20 µl) of CpGenome™ Human Non-Methylated DNA Standard or 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 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, and only the U primer set amplified the unmethylated Standard.View CpGenome™ Human Methylated & Non-Methylated DNA Standard Set on Millipore’s website