香蕉SSR反应体系的优化  

Optimization of Simple Sequence Repeat Reaction System in Banana (Musa sp.)

作　　者：郑丽珊[1] 王静毅[1] 冀小蕊[1] 武耀廷[1] 

机构地区：[1]中国热带农业科学院热带作物生物技术研究所,海南海口571101

出　　处：《热带农业科学》2007年第2期14-17,共4页Chinese Journal of Tropical Agriculture

基　　金：国家自然科学基金项目(No.30460070);海南省自然科学基金指导项目(No.80431)。

摘　　要：以香蕉品种AAcv Rose为试材,研究了香蕉SSR技术中PCR反应体系的主要成分对SSR扩增结果的影响,并比较了聚丙烯酰胺凝胶及琼脂糖凝胶电泳检测扩增产物多态性的差异。结果表明:在20μL的反应体系中,各成分的最适用量分别为TaqDNA酶0.5U/mL;Mg2+2.5mmol/L;引物0.4μmol/L;模板DNA20～40ng/μL;dNTPs0.1mmol/L。利用24个香蕉品种验证此反应体系,80g/L的非变性聚丙稀酰胺凝胶电泳检测结果显示,扩增产物在150～300bp之间,不同品种间DNA谱带具有多态性。聚丙稀酰胺凝胶电泳检测的DNA多态性高于琼脂糖。An attempt was made to study the effects of major elements in PCR reaction system on SSR analysis with banana AAcv Rose （Musa sp.） as the material, and compared was the difference between the single sequence repeat polymorphisms detected by non-denatured polyacrylamide and agar gel electrophoresis. The results were found optimal under the conditions of 0.5 U/mL Taq DNA polymerase, terminal concentration of 2.5 mmol/L MgCl2, 0.1 mmol/L dNTPs, 0.4 μmol/L primers and 20-40 ng/μL templates of DNA in total volume of 20 μL. To evaluate this SSR system, 24 accessions of banana were used. The 80 g/L polyacrylamide gel electrophoresis showed that the PCR products were between 100-300 bp and polymorphic in DNA among the banana accessions. Polyacrylamide gel had higher resolution than that of agar gel.

关 键 词：香蕉 简单重复序列 体系优化 

分 类 号：S668.1[农业科学—果树学]