About Human Methylated & Non-methylated DNA Set
The Human Methylated & Non-methylated DNA Set consists of two control DNAs (a methylated human DNA standard and a non-methylated human DNA standard) together with a set of specifically designed primers that can be used in conjunction with the EZ DNA Methylation™, EZ DNA Methylation-Gold™, and EZ DNA Methylation-Direct™ kits from Zymo Research to assess the efficiency of bisulfite-mediated conversion of DNA. The non-methylated human DNA is purified from the HCT116 DKO (double knock-out) cell line, which contains genetic knockouts of both DNA methyltransferases DNMT1 (-/-) and DNMT3b (-/-). The DNA derived from HCT116 DKO cells has a low level of DNA methylation and therefore can be used as a negative control for DNA methylation analysis (see below). The methylated human DNA standard is purified HCT116 DKO DNA that has been enzymatically methylated at all cytosine positions comprising CG dinucleotides by CpG Methylase and can be used as a positive control for DNA methylation analysis.
Figure 1. An assay for complete methylation by M.SssI methytransferase. Digestion of non-methylated and methylated HCT116 DKO DNA with restriction enzymes MspI and HpaII. MspI digests both nonmethylated and methylated DNA. HpaII is sensitive to CpG methylation.