DNA甲基化分析中重亚硫酸盐处理DNA转化效率的评估方法

doi:10.16288/j.yczz.15-129

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HEREDITAS(Beijing) ›› 2015, Vol. 37 ›› Issue (9): 939-944.doi: 10.16288/j.yczz.15-129

• Technique and Method • Previous Articles Next Articles

The method of estimating bisulfite conversion rate in DNA methylation analysis

Yangyang Liu¹, Hengmi Cui^{1, 2}

1. Jiangsu Province Key Lab of Molecular Medicine, School of Medicine, Nanjing University, Nanjing 210093, China;
2. Institute of Epigenetics & Epigenomics, Yangzhou University, Yangzhou 225009, China

Received:2015-03-26 Revised:2015-06-16 Online:2015-09-20 Published:2015-09-20

Abstract

Abstract: To establish an effective method to estimate the conversion rate of bisulfite-treated genomic DNA, TaqMan qPCR assay was performed using probes and primers that are specific for bisulfite-converted or -unconverted DNA standard samples separately. Then two linear standard curves were generated by plotting Ct values against logarithm of absolute DNA amount with serial dilutions of the bisulfite-converted or unconverted DNA samples. Based on two standard curves, the unknown bisulfite-treated genomic DNA sample was analyzed using the same TaqMan probes and the bisulfite conversion rate was precisely estimated. This method was further verified to be reliable using known mixed bisulfite-converted and -unconverted DNA templates as well as DNA samples treated with different bisulfite kits. These results showed that this method can effectively estimate bisulfite conversion rate of genomic DNA and thus provides a reliable and quick method for accurate analyses of DNA methylation.

Key words: DNA methylation, bisulfite, TaqMan qPCR, conversion rate

Yangyang Liu, Hengmi Cui. The method of estimating bisulfite conversion rate in DNA methylation analysis[J]. HEREDITAS(Beijing), 2015, 37(9): 939-944.

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