铁线莲属植物SSR-PCR反应体系的建立与优化  

Establishment and Optimization of SSR-PCR Reaction System of Clematis

作　　者：李勇慧 于相丽 周晓君 LI Yonghui;YU Xiangli;ZHOU Xiaojun

机构地区：[1]洛阳师范学院生命科学学院,河南洛阳471934

出　　处：《智慧农业导刊》2021年第21期32-35,共4页JOURNAL OF SMART AGRICULTURE

基　　金：国家自然科学基金面上项目(31870697)。

摘　　要：为对铁线莲进行反应体系的建立和优化,本实验采用了CTAB法提取铁线莲的DNA,然后以各条引物的Tm温度为参照,设置3个温度梯度,来筛选退火的最佳温度。同样设置浓度梯度筛选出DNA模板的最适浓度,最后用最佳反应体系和反应程序对100条引物进行筛选。测得最佳体系中加入0.5μL DNA,并且确定了各个引物的最佳退火温度,在此基础上选出10号、15号、30号、46号、96号和100号6个多态性良好的引物,为下一步的铁线莲种质遗传多样性分析打下基础。To establish and optimize the reaction system of clematis.The DNA of Clematis was extracted by CTAB method,and then with the Tm temperature of each primer as the reference,three temperature gradients were set to screen the best annealing temperature.The optimal concentration of DNA template was screened by the same concentration gradient.Finally,100 primers were screened by the best reaction system and reaction procedure.0.5μL DNA was added to the optimal system,and the best annealing temperature of each primer was determined.On this basis,six primers with good polymorphism were selected to lay a foundation for further genetic diversity analysis of Clematis germplasm.

关 键 词：铁线莲属 SSR 反应体系的建立与优化 引物筛选 

分 类 号：S68[农业科学—观赏园艺]