High-throughput method for analyzing methylation of CpGs in targeted genomic regions

Nautiyal S.; Carlton V.E.H.; Lu Y.Ireland J.S.Flaucher D.Moorhead M.Gray J.W.Spellman P.Mindrinos M.Berg P.Faham M.

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High-throughput method for analyzing methylation of CpGs in targeted genomic regions

机译：High-throughput method for analyzing methylation of CpGs in targeted genomic regions

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A unique microarray-based method for determining the extent of DNA methylation has been developed. It relies on a selective enrichment of the regions to be assayed by target amplification by capture and ligation (mTACL). The assay is quantitatively accurate, relatively precise, and lends itself to high-throughput determination using nanogram amounts of DNA. The measurements using mTACLs are highly reproducible and in excellent agreement with those obtained by sequencing (r = 0.94). In the present work, the methylation status of >145,000 CpGs from 5,472 promoters in 221 samples was measured. The methylation levels of nearby CpGs are correlated, but the correlation falls off dramatically over several hundred base pairs. In some instances, nearby CpGs have very different levels of methylation. Comparison of normal and tumor samples indicates that in tumors, the promoter regions of genes involved in differentiation and signaling are preferentially hypermethylated, whereas those of housekeeping genes remain hypomethylated. mTACL is a platform for profiling the state of methylation of a large number of CpG in many samples in a cost-effective fashion, and is capable of scaling to much larger numbers of CpGs than those collected here.

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《Proceedings of the National Academy of Sciences of the United States of America》 |2010年第28期|12587-12592|共6页
作者
Nautiyal S.; Carlton V.E.H.; Lu Y.Ireland J.S.Flaucher D.Moorhead M.Gray J.W.Spellman P.Mindrinos M.Berg P.Faham M.;
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作者单位

Affymetrix Inc., Santa Clara, CA 95051, United States;

Lawrence Berkeley National Laboratory, Berkeley, CA 94720, United States;

Stanford Genome Technology Center, Palo Alto, CA 94304, United StatesDepartment of Biochemistry, Stanford University, Stanford, CA 94305, United States;

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收录信息美国《科学引文索引》(SCI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
原文格式 PDF
正文语种英语
中图分类自然科学总论;
关键词
Array; Technology; Tumor;

High-throughput method for analyzing methylation of CpGs in targeted genomic regions

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