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Quantitative DNA methylation analysis by fluorescent polymerase chain reaction single‐strand conformation polymorphism using an automated DNA sequencer
Author(s) -
Suzuki Hiromu,
Itoh Fumio,
Toyota Minoru,
Kikuchi Takefumi,
Kakiuchi Hideki,
Hinoda Yuji,
Imai Kohzoh
Publication year - 2000
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/(sici)1522-2683(20000301)21:5<904::aid-elps904>3.0.co;2-4
Subject(s) - bisulfite , dna sequencer , sodium bisulfite , bisulfite sequencing , microbiology and biotechnology , single strand conformation polymorphism , dna methylation , methylation , polymerase chain reaction , illumina methylation assay , dna , methylated dna immunoprecipitation , biology , chemistry , dna sequencing , biochemistry , gene , gene expression , organic chemistry
A novel DNA methylation assay technique, termed bisulfite single‐strand conformation polymorphism (bisulfite‐SSCP), is a combination of sodium‐bisulfite modification and fluorescence‐based polymerase chain reaction (PCR)‐SSCP. After bisulfite treatment followed by PCR amplification, methylated and unmethylated alleles can be simultaneously separated in a nondenaturing gel using an automated DNA sequencer. Using bisulfite‐SSCP, methylation of hMLH1 was detected in a quantitative manner. This method is not only simple, quick, accurate, and quantitative, but detailed information about methylation is also available with less work. Methylation analysis of large numbers of samples for multiple loci will be facilitated by bisulfite‐SSCP.