[Application of created restriction site PCR-RFLP to identify alcohol dehydrogenase 2 gene polymorphism]
1Department of Occupational Health, School of Public Health, Fudan University, Laboratory of Public Health Safety, Ministry of Education, Shanghai 200032, China. jiaojie66@126.com
Objective:
To develop a appropriate assay for identifying single nucleotide polymorphism (SNP) of alcohol dehydrogenase 2 (ADH2) gene.
Methods:
According to base substitution situation of one single base mutational site, we designed the present study primers. One of the primers was designed on the basis of neighbourhood sequence of the mutational site, that is, we made one mismatch base to let product a new enzyme site between the 3' end of the primer and the single base mutation type after the PCR amplification. Then PCR-RFLP was adopted to identify the SNP in ADH2 gene.
Results:
One primer pair can get target products containing ADH2 SNP site by PCR, restriction enzymes Bsh1236I were adopted to identify the SNP site. The expected results were reached.
Conclusion:
It suggested that the method of detecting the SNP of ADH2 based on CRS-PCR-RFLP theory is facilitated, economic, and rapid.
