都支杜鹃ISSR扩增条件的优化  

Optimization of ISSR Amplification Conditions for Rhododendron Shanii

作　　者：颜亮[1] 赵凯[2] 

机构地区：[1]皖南医学院药学系,安徽芜湖241000 [2]安庆师范学院生命科学学院,安徽安庆246011

出　　处：《绵阳师范学院学报》2011年第11期87-90,共4页Journal of Mianyang Teachers＇ College

摘　　要：对珍稀濒危物种都支杜鹃的ISSR扩增体系中的Mg2+浓度、dNTP浓度、Taq酶含量和底物含量进行优化,并在优化后的基础上筛选出8条效果较好的引物,在梯度PCR仪中设置温度梯度,找到这些引物的最佳退火温度。优化结果表明,至少在一定的范围内底物浓度和Taq酶含量对PCR反应结果影响不大,相对而言,dNTP浓度和Mg2+浓度对反应结果的影响更加显著。优化后的15 ul PCR反应体系中包括20 ng模板DNA、0.3μM引物、1.5 ul Buffer(Mg2+free),0.5 U Taq酶、1.5 mM MgCl2和0.1 mM dNTP。Factors that affect ISSR PCR amplification, such as Mg2 ＋ concentration, dNTP concentration, were optimized by using the genomic DNA of a rare and endangered species Rhododendron shanii as material, on the foundation of optimization, 8 primers were selected. The optimal annealing temperature of every primer was selected by using gradient PCR instrument. The optimized result indicates that at least in certain scope template DNA dosage and Taq DNA polymerase affect little on the result of PCR reaction, relatively, Mg^2 ＋ concentration and dNTP concentration have significant effects to the result, The suitable 15ul PCR reaction should contain 20ng template DNA, 0.3μM primer, 1.5ul buffer （ Mg2 ＋ free） 0.5U Taq DNA polymerase, 1.5 mM MgCl2 and 0.1 mM dNTP.

关 键 词：都支杜鹃 ISSR PCR扩增 条件优化 

分 类 号：Q943[生物学—植物学]