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.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Xiong Z,Laird PWdoi
10.1093/nar/25.12.2532subject
Has Abstractpub_date
1997-06-15 00:00:00pages
2532-4issue
12eissn
0305-1048issn
1362-4962pii
gka376journal_volume
25pub_type
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