COBRA: a sensitive and quantitative DNA methylation assay | Zendy

Zhongguo Xiong | Zendy; Peter W. Laird | Zendy

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COBRA: a sensitive and quantitative DNA methylation assay

Author(s) -

Zhongguo Xiong,

Peter W. Laird

Publication year - 1997

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/25.12.2532

Subject(s) - biology , dna methylation , cobra , dna , microbiology and biotechnology , sodium bisulfite , methylation , illumina methylation assay , genomic dna , bisulfite sequencing , methylated dna immunoprecipitation , restriction enzyme , amplified fragment length polymorphism , polymerase chain reaction , bisulfite , gene , genetics , gene expression , chemistry , population , demography , organic chemistry , sociology , computer science , genetic diversity , programming language

We report here on a quantitative technique called COBRA to determine DNA methylation levels at specific gene loci in small amounts of genomic DNA. Restriction enzyme digestion is used to reveal methylation-dependent sequence differences in PCR products of sodium bisulfite-treated DNA as described previously. We show that methylation levels in the original DNA sample are represented by the relative amounts of digested and undigested PCR product in a linearly quantitative fashion across a wide spectrum of DNA methylation levels. In addition, we show that this technique can be reliably applied to DNA obtained from microdissected paraffin-embedded tissue samples. COBRA thus combines the powerful features of ease of use, quantitative accuracy, and compatibility with paraffin sections.

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