[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.


CONCLUSIONS
The methods for four SNPs of MGMT determinated by CRS-PCR-RFLP theory could be facility, economy, and rapidness.
Wang
Wei
W
Department of Occupational Health, School of Public Health, Fudan University, Laboratory of Public Health Safety, Ministry of Education, Shanghai 200032, China. ww375@126.com
Miao
Wenbin
W
Qiu
Yulan
Y
Xia
Zhaolin
Z
D002617:Chemical Industry; D004247:DNA / Q000097:blood; D015254:DNA Modification Methylases / Q000235:genetics*; D045643:DNA Repair Enzymes / Q000235:genetics*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D020641:Polymorphism, Single Nucleotide; D015183:Restriction Mapping; D025521:Tumor Suppressor Proteins / Q000235:genetics*
D025521:Tumor Suppressor Proteins; D004247:DNA; D015254:DNA Modification Methylases; C515491:MGMT protein, human; D045643:DNA Repair Enzymes