贵州山核桃ISSR-PCR反应体系建立及优化  

Establishment and Optimization of ISSR-PCR Reaction System of Carya Cathayensis

作　　者：邱国俊[1] 朱娅 张培[1] 苗贵东[1] 郭计华[1] QIU Guojun;ZHU Ya;ZHANG Pei;MIAO Guidong;GUO Jihua(Xingyi Normal University for Nationalities,Xingyi,Guizhou 562400)

机构地区：[1]兴义民族师范学院,贵州兴义562400

出　　处：《兴义民族师范学院学报》2020年第2期114-119,共6页Journal of Minzu Normal University of Xingyi

基　　金：兴义民族师范学院教授基金项目,项目名称:贵州山核桃种质资源遗传多样性研究(17XYJS04);贵州省教育厅青年科技人才成长项目,项目名称:贵州核桃微卫生(SSR)单核甘酸多态性(SNP)分子记开发及应用研究(黔教合KY〔2016〕327号)。

摘　　要：为建立贵州山核桃ISSR-PCR最佳反应体系,采用单因素试验的方法,对影响ISSR-PCR反应体系的模板DNA浓度、MgCl2浓度、d NTPs浓度、Taq酶用量、引物浓度等各因素进行了研究,确立了贵州山核桃ISSR-PCR的最佳反应体系。结果显示:最优反应体系的模板DNA浓度为40ng/u L、引物浓度为0.3umol/L、Mg2+的浓度2.9mmol/L、酶浓度0.3U、d NTPs浓度0.2mmol/L,退火温度为54℃。建立该反应体系可为贵州山核桃ISSR标记开发、物种分类鉴定、遗传图谱构建等后续分析奠定基础。in order to establish the optimal issr-pcr reaction system of guizhou pecan, this study adopted the single-factor test method to optimize the factors affecting issr-pcr reaction system, such as template DNA concentration, MgCl2 concentration, dNTPs concentration, Taq enzyme dosage, primer concentration and so on, and established the optimal issr-pcr reaction system of guizhou pecan. The results showed that the template concentration of the optimal reaction system was 40 ng/uL,the primer concentration was0.3 mmol/L, the concentration of Mg2+ was 2.9 mmol/L,the enzyme concentration was 0.3 U,and the dNTPs concentration was0.2 mmol/L. The establishment of this reaction system can lay a foundation for ISSR marker development, species classification and identification,genetic map construction and other subsequent analysis of guizhou pecan.

关 键 词：贵州山核桃 ISSR-PCR反应体系 单因素优化 

分 类 号：Q94-3[生物学—植物学]