[Application of double created restriction site PCR-RFLP to identify MGMT gene polymorphisms].

OBJECTIVE: To develop a proper assay for identifying single nucleotide polymorphisms( SNPs) of the MGMT gene. METHODS: PCR primers were designed by create restriction site (CRS) method, then polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was adopted to identify four SNPs in MGMT gene. RESULTS: By PCR, one primer pair yielded target products containing MGMT84 SNP site, and the other primer pair yielded target products containing MGMT143, 160, 178 SNP sites. Four restriction enzymes were adopted to identify the four SNPs, respectively. The effects of PCR and RFLP were good. CONCLUSION: The methods for four SNPs of MGMT determinated by CRS-PCR-RFLP theory could be facility, economy, and rapidness.