鳜SSR-PCR反应条件的优化探索  

Discussion about the Reaction Conditions of Microsatellite PCR in Siniperca chuatsi

作　　者：匡刚桥[1] 吴青[1] 郑曙明[1] 陈建[1] 鲁双庆[2] 

机构地区：[1]西南大学水产系,重庆荣昌402460 [2]长沙大学生物工程与环境科学系,湖南长沙410003

出　　处：《水生态学杂志》2009年第3期128-130,共3页Journal of Hydroecology

基　　金：西南大学荣昌校区科研项目(2007s08)

摘　　要：以鳜(Siniperca chuatsi)基因组DNA为试验材料,研究MgCl2浓度、dNTPs浓度、TaqDNA聚合酶3个反应参数对微卫星PCR的影响,建立一套适合鳜微卫星PCR反应的最佳体系。结果表明,在25μL的反应体系中,Mg2+、dNTPs、TaqDNA聚合酶3个参数的适宜浓度分别为1.2mmol/L、0.2μmol/L和0.04U/μL。PCR反应程序为94℃变性5min,94℃30s,56℃1min,72℃1min,共30个循环,最后72℃延伸5min。In this study, A microsatellite DNA PCR reaction system of Siniperca chuatsi was constructed by the optimization of the concentrations of Mg2＋ ,dNTPS and Taq DNA Polymerase. The results showed that： The optimalized PCR system with total volume 25 μL including MgCl2 at 1.2 mmoL/L, dNTP at 0.2 μmol/L, Taq DNA polymerase at 0.04 U/μL. The PCR reaction program with 5 minutes at 94 ℃ ,which is followed by 30 cycles ,each with 30 seconds at 94℃, 1 minute at 56 ℃, 1 minute at 72℃, and a final extension at 72 ℃ for 5 minutes.

关 键 词：鳜 微卫星 反应条件 

分 类 号：S948[农业科学—水产养殖]