![]() ![]() Bisulfite conversion of cytosines requires a single-strand target, so the process is typically carried out at elevated temperature. ![]() ![]() First, the procedure induces widespread damage to DNA in general. While bisulfite treatment is the gold standard for DNA epigenetic analysis, it has two significant drawbacks. For example, physiologically, 5mC has been shown to occur primarily in symmetric CpG dinucleotides in mammalian genomes 4, where it plays an important role in the regulation of gene expression 5 and has consequently been implicated in a variety of diseases 6 including cancer 7. In combination with the growing availability of sequencing technologies, this simple treatment has enabled a large number of studies that have been able to determine the genomic positions of 5mC as well as highlight its importance in diverse biological processes. It is the most widely studied DNA base variant, largely because of the early advent of a technique with which it could be probed it was demonstrated 2, 3 as early as 1970 that exposure to sodium bisulfite is capable of deaminating cytosines and converting them to uracils, but that this chemical reaction is blocked by methylation. Composed structurally of a cytosine nucleobase with a methyl group at the fifth carbon atom, the epigenetic modification 5-methylcytosine (5mC) has an overall prevalence of ~ 4% (5mC/C) in the human genome 1. ![]()
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