Genome-scale DNA methylation mapping of clinical samples at single-nucleotide resolution

被引：224
作者：
Gu, Hongcang [1 ]
Bock, Christoph [1 ,2 ,3 ,4 ]
Mikkelsen, Tarjei S. [1 ]
Jaeger, Natalie [1 ,2 ,3 ]
Smith, Zachary D. [1 ,2 ,3 ]
Tomazou, Eleni [1 ,2 ,3 ]
Gnirke, Andreas [1 ]
Lander, Eric S. [1 ,5 ,6 ]
Meissner, Alexander [1 ,2 ,3 ]
机构：
[1] Broad Inst, Cambridge, MA USA
[2] Harvard Univ, Dept Stem Cell & Regenerat Biol, Cambridge, MA 02138 USA
[3] Harvard Stem Cell Inst, Cambridge, MA USA
[4] Max Planck Inst Informat, Saarbrucken, Germany
[5] MIT, Dept Biol, Cambridge, MA USA
[6] Harvard Univ, Sch Med, Dept Syst Biol, Boston, MA USA
来源：
NATURE METHODS | 2010年 / 7卷 / 02期
基金：
美国国家卫生研究院;
关键词：
COLORECTAL-CANCER; MAPS; EPIGENOME; COLON;
D O I：
10.1038/nmeth.1414
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
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.
引用
收藏
页码：133 / U69
页数：6
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