Skip to main page content
Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
Comparative Study
, 25 (12), 2529-31

Rapid Quantitation of Methylation Differences at Specific Sites Using Methylation-Sensitive Single Nucleotide Primer Extension (Ms-SNuPE)

Affiliations
Comparative Study

Rapid Quantitation of Methylation Differences at Specific Sites Using Methylation-Sensitive Single Nucleotide Primer Extension (Ms-SNuPE)

M L Gonzalgo et al. Nucleic Acids Res.

Abstract

We have developed a rapid quantitative method (Ms-SNuPE) for assessing methylation differences at specific CpG sites based on bisulfite treatment of DNA followed by single nucleotide primer extension. Genomic DNA was first reacted with sodium bisulfite to convert unmethylated cytosine to uracil while leaving 5-methylcytosine unchanged. Amplification of the desired target sequence was then performed using PCR primers specific for bisulfite-converted DNA and the resulting product isolated and used as a template for methylation analysis at the CpG site(s) of interest. This methylation-sensitive technique has several advantages over existing methods used for detection of methylation changes because small amounts of DNA can be analyzed including microdissected pathology sections and it avoids utilization of restriction enzymes for determining the methylation status at CpG sites.

Similar articles

See all similar articles

Cited by 78 PubMed Central articles

See all "Cited by" articles

References

    1. Nucleic Acids Res. 1990 Feb 11;18(3):687 - PubMed
    1. Proc Natl Acad Sci U S A. 1991 Feb 15;88(4):1143-7 - PubMed
    1. Proc Natl Acad Sci U S A. 1992 Mar 1;89(5):1827-31 - PubMed
    1. PCR Methods Appl. 1992 Feb;1(3):160-3 - PubMed
    1. Nucleic Acids Res. 1997 Jun 15;25(12):2532-4 - PubMed

Publication types

LinkOut - more resources

Feedback