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Nucleic Acids Res. 1997 Jun 15;25(12):2532-4.

COBRA: a sensitive and quantitative DNA methylation assay.

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  • 1Department of Surgery, University of Southern California School of Medicine, The Norris Comprehensive Cancer Center, Room 6415, Mailstop #73, 1441 Eastlake Avenue, Los Angeles, CA 90033, USA.

Abstract

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.

PMID:
9171110
[PubMed - indexed for MEDLINE]
PMCID:
PMC146738
Free PMC Article
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