亚硫酸氢盐
脱氨基
亚硫酸氢钠
亚硫酸氢盐测序
胞嘧啶
甲基化DNA免疫沉淀
化学
DNA甲基化
DNA
5-甲基胞嘧啶
基因组DNA
生物化学
分子生物学
计算生物学
基因
生物
有机化学
酶
基因表达
作者
Hikoya Hayatsu,Kazuo Negishi,Masahiko Shiraishi,Kaoru Tsuji,K. Moriyama
出处
期刊:Nucleic Acids Symposium Series
[Oxford University Press]
日期:2007-11-01
卷期号:51 (1): 47-48
被引量:3
摘要
Methylation at position 5 of cytosine in DNA plays a major role in epigenetic gene control. The methylation analysis can be performed by bisulfite genomic sequencing. Conventional procedures in this analysis include a treatment of single stranded DNA with 3-5 M sodium bisulfite at pH 5 and at 50-55 degrees for 4-20 hr. This will convert cytosine into uracil, while 5-methylcytosine resists this deamination. Amplification by PCR of the bisulfite-treated DNA followed by sequencing reveals the positions of 5-methylcytosine in the gene. We reported recently that the whole procedure can be speeded up by use of a highly concentrated bisulfite solution, 10 M ammonium bisulfite. We also reported that urea, which has been often added to the reaction mixture with the purpose of facilitating the reaction, may not work as anticipated. This time, we would like to address the need for further investigating the chemistry of the bisulfite modification of DNA. Particularly important is to study side reactions that may occur due to the exhaustive bisulfite treatment required for achieving complete deamination of all the cytosine residues in a given sample of DNA.
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