小型西瓜SRAP技术体系优化  

Optimization of SRAP Reaction System on Mini-watermelon

作　　者：江海坤[1] 王明霞[1] 严从生[1] 王艳[1] 马绍鋆[1] 董言香[1] 方凌[1] 张其安[1] 

机构地区：[1]安徽省农业科学院园艺研究所,合肥230031

出　　处：《中国农学通报》2011年第28期240-244,共5页Chinese Agricultural Science Bulletin

基　　金：安徽省自然科学基金项目"西瓜裂果性状QTL定位"(10040606Q29);国家西甜瓜产业技术体系合肥综合试验站(CARS-26-31)

摘　　要：为探讨小型西瓜种质遗传分析奠定基础以及不类型西瓜SRAP技术体系的通用性,以小型西瓜F'1秀丽'为试材,利用正交试验设计,对SRAP-PCR反应体系中的Mg2+浓度、dNTPs浓度、引物浓度、Taq聚合酶浓度和模板DNA浓度进行5因素4水平的筛选分析,用Me3-Em3引物组合进行PCR扩增以确定最优反应体系;进一步应用该优化反应体系,对5个不同引物和37份不同果型西瓜资源DNA进行SRAP-PCR扩增。结果表明,小型西瓜SRAP-PCR最佳反应体系为:10×PCR buffer 2μL、Mg2+3.0mmol/L,dNTPs0.2mmol/L,引物0.5μmol/L,模板DNA 40ng、Taq聚合酶0.5U,总体积为10μL。不同果型西瓜资源DNA进行SRAP-PCR扩增,电泳条带清晰、稳定性好,说明不同果型西瓜种质SPAP体系具有通用性。To establish SRAP reaction system for mini-watermelon and explore the generality of the SRAP technology system among different type of watermelon resources,the SRAP reaction system was optimized,in terms of Mg 2＋,dNTPs,primers,Taq polymerase and template DNA 5 factors and their 4 levels with the primer Em3-Me3,by an orthogonal design.And further application of the optimal reaction system was applied among different type of watermelon resources.The results indicated that the watermelon appropriate SRAP-PCR reaction system in a 10 μL reaction system was：10× PCR buffer 2 μL,Mg 2＋ 3.0 mmol/L,dNTPs 0.2 mmol/L,primer 0.5 μmol/L,template DNA 40 ng,Taq polymerase 0.5 U.37 mini-watermelon germplasms were used to test the stabilization of the optimized reaction system.The optimized reaction system was very stable and can well meet the different type watermelon requirements of genomic DNA amplification.

关 键 词：小型西瓜 SRAP 正交 体系优化 

分 类 号：S651[农业科学—果树学]