番茄微卫星标记(SSR)反应体系的优化研究  

Optimization of SSR Analysis System in Tomato

作　　者：刘杨[1] 陈火英[2] 魏毓棠[1] 

机构地区：[1]沈阳农业大学园艺学院,沈阳110161 [2]上海交通大学植物科学系,上海201101

出　　处：《沈阳农业大学学报》2005年第2期155-158,共4页Journal of Shenyang Agricultural University

基　　金："十五"国家"863"计划基金项目(2001AA67010)

摘　　要：利用番茄材料研究了番茄SSR技术中PCR体系的主要成分对SSR扩增结果的影响,并比较了采用聚丙烯酰胺凝胶和琼脂糖电泳检测扩增产物多态性的差异。结果表明:番茄SSR的反应程序为94℃预变性2min;94℃变性30s,50℃复性30s,68℃延伸30s,35个循环;最后72℃延伸8min为佳。反应体系中,模板DNA的适宜浓度为20～40ng,适宜的引物浓度为0.4μmol·L-1,dNTP的适宜浓度为200μmol·L-1,Mg2+的最适浓度范围为2.0mmol·L-1,Taq聚合酶在15μL反应体系中宜加入0.65U。PCR产物检测时聚丙烯酰胺凝胶的分辨率显著高于琼脂糖电泳。The SSR amplified results of tomato and comparative analysis of polymorphism of SSR detected on two gel electrophoresis systems were studied. A standard SSR system fitting for tomato was established. The results showed that the optimized PCR cycles started at 94℃（2min）, followed by 35 cycles of 30s at 94℃,30s at 50℃ and 30s at 68℃, and then extended 8min at 72℃. The optimized amplification conditions were. 20-40 ng DNA, 0.4 μmol· L^-1 of each primer, 200μmol·L^-1 of dNTP, 2.0mmol· L^-1 Mg^2＋ and 0.65 unit Amplitaq DNA polymerse in 15μL volume. 6% Polyacrylamide gel had higher ability than 3% agarose gel in discriminating polymorphism.

关 键 词：番茄·微卫星标记 反应体系 

分 类 号：S641.2[农业科学—蔬菜学]