Broad Institute, Cambridge, Massachusetts, USA.
Nat Methods. 2010 Feb;7(2):133-6. doi: 10.1038/nmeth.1414. Epub 2010 Jan 10.
Bisulfite sequencing measures absolute levels of DNA methylation at single-nucleotide resolution, providing a robust platform for molecular diagnostics. We optimized bisulfite sequencing for genome-scale analysis of clinical samples: here we outline how restriction digestion targets bisulfite sequencing to hotspots of epigenetic regulation and describe a statistical method for assessing significance of altered DNA methylation patterns. Thirty nanograms of DNA was sufficient for genome-scale analysis and our protocol worked well on formalin-fixed, paraffin-embedded samples.
亚硫酸氢盐测序以单核苷酸分辨率测量 DNA 甲基化的绝对水平，为分子诊断提供了一个强大的平台。我们针对临床样本的全基因组分析对亚硫酸氢盐测序进行了优化：在此，我们概述了如何通过限制性消化将亚硫酸氢盐测序靶向到表观遗传调控的热点，并描述了一种用于评估 DNA 甲基化模式改变的显著性的统计方法。30 纳克的 DNA 足以进行全基因组分析，并且我们的方案在福尔马林固定、石蜡包埋的样本上效果良好。
