仙茅属植物SRAP-PCR反应体系的优化  

Optimization of SRAP-PCR System for Curculigo L.Plant

作　　者：陈大霞[1] 李隆云[1] 李泉森[1] 秦松云[1] 钟国跃[1] 

机构地区：[1]重庆市中药研究院,重庆400065

出　　处：《分子植物育种》2007年第F11期191-195,共5页Molecular Plant Breeding

基　　金：重庆市科委攻关项目(6455)

摘　　要：通过单因子和双因子实验对仙茅属植物SRAP-PCR反应体系中主要成分(Mg^(2+),dNTP、引物、模板和Taq DNA聚合酶)进行优化,并比较了琼脂糖凝胶电泳与非变性PAGE电泳的检测效果。建立了适合仙茅属植物SRAP分析的反应体系:25μL体系中内含1×PCR buffer、2.0 mmol/L Mg^(2+)、250μmol/L dNTP、0.2μmol/L引物、40ng模板、1 U Taq酶。非变性聚丙烯酰胺凝胶电泳分辨率较高且带型清晰,能更准确反映仙茅属植物间的差异。优化的反应体系可以用于仙茅属植物的SRAP分析。In this research SRAP reaction conditions were optimized by single or dual factor experiment that involved in the main reaction components, i.e. the concentration of Mg^2＋, dNTP, primer, and template DNA, as well as Taq DNA polymerase dosage. The products of SRAP were identified both by agarose gel and non-denaturing polyacrylamide gel electrophosresis. It was obvious that the optimized reaction system would be suitable for PCR ananlysis of Curculigo plant, which might be the 25 μL amplification reaction system containing 1 ×PCR buffer, 2.0 mmol/L Mg^2＋, 250 μmol/L dNTP, 0.2 μmol/L primer, 40 ng template DNA, and 1 U Taq DNA polymerase. And also it is worthy mentioned that the non-denaturing PAGE approach would be more productive and informative for SRAP-PCR, which could obtain the high resolution banding patterns.

关 键 词：仙茅属 SRAP PCR反应体系 优化 

分 类 号：Q943[生物学—植物学]