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PMID: 7785768 Published · ppublish English Journal Article

A bisulfite method of 5-methylcytosine mapping that minimizes template degradation.

Analytical biochemistry ·Vol. 226 ·No. 1 ·1995-03-20 ·Pages 161-6

Raizis AM, Schmitt F, Jost JP

Abstract

The bisulfite method is a highly sensitive approach to 5-methylcytosine mapping that utilizes the capability of the polymerase chain reaction to exponentially amplify DNA. We have observed that the bisulfite reaction results in a significant level of template degradation due to DNA depurination. Furthermore, our data suggest that the DNA fragmentation which occurs limits the sensitivity of the method. We describe a simple solution to limit degradation of the DNA template.

MeSH Terms
5-Methylcytosine Base Sequence Cytosine/analogs & derivatives,analysis DNA/chemistry DNA Modification Methylases/metabolism DNA Primers Hydrogen-Ion Concentration Methylation Molecular Sequence Data Polymerase Chain Reaction Sequence Analysis, DNA Sulfites/pharmacology Templates, Genetic
Chemicals
DNA Primers Sulfites 5-Methylcytosine Cytosine DNA DNA Modification Methylases hydrogen sulfite
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Raizis A M
Friedrich Miescher Institut, Basel, Switzerland.
Schmitt F
Jost J P
Article Info
Journal
Analytical biochemistry
Abbr.
Anal Biochem
ISSN
0003-2697
Published
1995-03-20
Pages
161-6
Language
English
Region
United States
NLM ID
0370535
Subset
IM
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