Summary: The bisulfite reaction reaction was first described in early 1970s and was used by Frommer et al to distinguish between cytosine and 5mCytosine (5mC) in DNA. In this reaction, DNA is first treated with sodium bisulfite to convert cytosine residues to uracil in single stranded DNA, under conditions whereby 5mC remains essentially non-reactive. The DNA sequence under investigation is then amplified by PCR with primers specific for bisulfite modified DNA. Many methods based on the same principle have been developed including bisulfite genomic sequencing PCR (BSP), methylation-specific PCR (MSP), COBRA, and methylation-sensitive single nucleotide primer extension (MS-SNuPE), with the first two being the most commonly used. All methods share the same procedure of modifying DNA with sodium bisulfite as the first step and subsequently PCR amplification with primers specific for modified DNA.
Bisulfite-PCR (for restriction and/or sequencing)
(Issa Lab) Bisulfite-PCR followed by restriction is a rapid and semi-quantitative method of analyzing DNA methylation. The PCR products are also suitable for either direct sequencing or cloning and sequencing. The most important step here is primer selection http://www.mdanderson.org/departments/methylation/... Added: Tue May 14 2002, Hits: 3033, Reviews: 0Write reviewCached
Singel Nucleotide Primer Extension (SNuPE)
(Methods.info) Single Nucleotide Primer Extension is a powerful method which can be used for the precise analysis of methylation in a certain position. http://www.methods.info/Methods/DNA_methylation/SN... Added: Sat Dec 20 2003, Hits: 625, Reviews: 0Write reviewCached