Quantitative DNA methylation analysis by fluorescent polymerase chain reaction single-strand conformation polymorphism using an automated DNA sequencer

✍ Scribed by Hiromu Suzuki; Fumio Itoh; Minoru Toyota; Takefumi Kikuchi; Hideki Kakiuchi; Yuji Hinoda; Kohzoh Imai

• Publisher: John Wiley and Sons
• Year: 2000
• Tongue: English
• Weight: 129 KB
• Volume: 21
• Category: Article
• ISSN: 0173-0835
• DOI: 10.1002/(sici)1522-2683(20000301)21:5<904::aid-elps904>3.0.co;2-4

✦ Synopsis

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.