贵州山核桃ISSR-PCR反应体系建立及优化

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。

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

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.