Bisulfite modification for analysis of DNA methylation.
Bisulfite is known to deaminate cytosine in nucleic acids, while 5-methylcytosine resists this bisulfite action. For this reason, bisulfite treatment has been used for detecting 5-methylcytosine in DNA, a minor component of eukaryotic DNA, presently recognized as playing an important role in the control of gene function. This procedure, called bisulfite genomic sequencing, is a principal method for the analysis of DNA methylation in various biological phenomena, including human diseases such as cancer. This unit describes an efficient procedure utilizing a newly developed high-concentration bisulfite solution. Protocols for this methodology are supplemented with discussions focused on chemical aspects of the bisulfite treatment.
Faculty of Pharmaceutical Sciences, Okayama University, Tsushima, Okayama, Japan.
Hikoya
H
Hayatsu
Masahiko
M
Shiraishi
Kazuo
K
Negishi
D044503:5-Methylcytosine / Q000737:chemistry; D000818:Animals; D001482:Base Composition; D001483:Base Sequence; D004247:DNA; D019175:DNA Methylation; D003641:Deamination; D016678:Genome / Q000235:genetics; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA / Q000379:methods*; D012995:Solubility; D012996:Solutions; D013447:Sulfites / Q000737:chemistry*
D012996:Solutions; D013447:Sulfites; D044503:5-Methylcytosine; D004247:DNA; C042345:hydrogen sulfite