重庆医学2012，Vol.41Issue(8)：764-765,768,封2,4.DOI:10.3969/j.issn.1671-8348.2012.08.014
碱裂解法快速提取口腔拭子DNA对CHRNA3基因多态性的研究
Rapid DNA extraction from buccal swabs by alkaline lysis method for study of CHRNA3 gene polymorphism
摘要
Abstract
Objective To establish a rapid method for DNA extraction from buccal swabs and investigate its application in analysis of cholinergic receptor nicotinic alpha3 (CHRNA3) gene polymorphism. Methods Lysis solution was made from NaOH and ethylenediaminetetraacetic acid(KDTA) ,and tris(hydroxymethyl)aminomethane-KDTA(TE) was served as neutralizing solution. DNA was extracted from buccal swabs via two steps:heating and neutralizing. Rs64953909 in CHRNA3 gene was genotyped by poly-merase chain reaction-restriction fragment length polymorphism(PCR-RFLP) assay using templates of extracted DNA. The samples of different genotypes were confirmed by direct sequencing analysis of PCR products. Results The target DNA bands obtained by PCR amplification and enzyme digestion were clear, and no nonspecific band was detected. The results of PCR-RFLP agreed well with the results of direct sequencing. Conclusion The alkaline lysis method for preparation of DNA from buccal swabs is rapid, simple, economical,reliable,and can be used for analysis of CHRNA3 gene polymorphism.关键词
多态性,限制性片段长度/序列分析,DNA/口腔拭子/尼古丁乙酰胆碱受体α3/碱裂解Key words
polymorphism,restriction fragment length/ sequence analysis, DNA/ buccal swab/ cholinergic receptor nicotinic alpha3/ alkaline lysis引用本文复制引用
朱伟锋,刘卓琦,吴金兰,余乐涵,万福生..碱裂解法快速提取口腔拭子DNA对CHRNA3基因多态性的研究[J].重庆医学,2012,41(8):764-765,768,封2,4.基金项目
江西省科技支撑计划资助项目(2009BSB09502). （2009BSB09502）
