苹果黑星病菌SSR反应体系的优化  

The Optimization of the SSR Reaction System for Venturia inaequalis

作　　者：董艳玲[1] 胡小平[1] 杨家荣[1] 苟建军[1] 

机构地区：[1]西北农林科技大学植保资源与病虫害治理教育部重点实验室,陕西杨陵712100

出　　处：《西北植物学报》2005年第6期1148-1152,共5页Acta Botanica Boreali-Occidentalia Sinica

基　　金：欧盟国际科技合作项目资助 ICA4-CT-2 0 0 1-10 0 0 1

摘　　要：通过对苹果黑星病菌基因组DNA的SSR反应中一些重要参数进行优化,结果表明,最适反应体系为:2 5μL体系中,10×Buffer Mg Cl2 2 0 mm ol/ L 2 .5μL ,d NTP 10 0μmol·L- 1 、引物0 .5μmol·L- 1 、Taq DNA聚合酶1.5 U ,DNA模板2 m g·L- 1 ,dd H2 O 19.5μL .PCR扩增程序为93℃,2 min,5 7℃,30 s,1个循环;72℃,1m in,93℃,30 s,5 7℃,30 s,4 0个循环;72℃,10 min.Some important parameters of the SSR reaction system aimed at genomic DNA of Venturia inaequalis were optimized, which demonstrated that the optimal reaction system was as follows: the 25 μL reaction system contained 2.5 μL 10×Buffer(25 mmol·L -1 MgCl_2), dNTP at 100 μmol·L -1 , primer at 0.5 μmol·L -1 , 1.5 U Taq DNA ploymerase, DNA template at 2 mg·L -1 , and 19.5 μL dd H_2O. The procedure of PCR amplification was as follows: PCR amplification was run at 93℃ for 2 minutes and then at 57℃ for 30 seconds and only 1 cycle was conducted ; the PCR amplification was run at 72 ℃ for 1 minute , then at 93℃ for 30 seconds and finally at 57 ℃ for 30 seconds , and 40 cycles were conducted; the PCR amplification was run at 72℃ for 10 minutes.

关 键 词：反应体系 黑星病菌 SSR 优化 苹果 基因组DNA DNA聚合酶 DNA模板 扩增程序 MOL TAQ PCR 循环 引物 

分 类 号：Q781[生物学—分子生物学] TQ323.8[化学工程—合成树脂塑料工业]