COBRA: a sensitive and quantitative DNA methylation assay
Open Access
- 1 June 1997
- journal article
- research article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 25 (12) , 2532-2534
- https://doi.org/10.1093/nar/25.12.2532
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 methylationdependent 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.Keywords
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