Brand：Sigma-Aldrich
Principles of the Technique Use of either the CpGenome DNA Modification Kit (Cat. No. S7820) or the CpGENOME Fast DNA Modification Kit (Cat. No. S7824) facilitates the initial bisulfite reactions, while the CpG WIZ GSTpi Amplification Kit contains the reagents required for the gene-specific PCR amplification reactions. Methylation of cytosines located 5′ to guanosine is known to have a profound effect on the expression of several eukaryotic genes (Bird, 1992). In normal cells, methylation occurs predominantly in CG-poor regions, while CG-rich areas, called CpG islands, remain unmethylated. Aberrant methylation of normally unmethylated CpG islands has been documented as a relatively frequent event in immortalized and transformed cells (Antequera, 1990) and has been associated with transcriptional inactivation of defined tumor suppressor genes in human cancers (Herman, 1994; Merlo, 1995). Glutathione S-Transferase (GSTpi) is an example of a gene that exhibits characteristic hypermethylation. Methylation-specific PCR (MSP) is a new technology for sensitive detection of abnormal gene methylation utilizing small amounts of DNA (Herman, 1996 ). This process employs an initial bisulfite reaction to modify the DNA, followed by PCR amplification with specific primers designed to distinguish methylated from unmethylated DNA.
