对节白蜡ISSR-PCR反应体系的建立与优化  

Establishment and Optimization of Fraxinus hupehensis ISSR-PCR Reaction System

作　　者：程建如 周明芹[1] 

机构地区：[1]长江大学园艺园林学院,湖北荆州434025

出　　处：《湖北农业科学》2016年第22期5962-5965,共4页Hubei Agricultural Sciences

基　　金：国家自然科学基金项目(31200528)

摘　　要：以对节白蜡(Fraxinus hupehensis Chiú,Shang et Su)基因组DNA为材料,采用正交试验和单因子试验对其ISSR-PCR反应体系进行了构建与优化。结果表明,在15μL反应体系中含DNA模版40 ng、引物终浓度0.7μmol/L、7.5μL 2×Taq PCR Plus Master Mix的扩增效果最好。该反应体系筛选出了14条稳定性强、多态性高、扩增效果较好的ISSR引物,并通过梯度PCR试验比较引物在不同退火温度下的扩增效果,从而确定引物最佳退火温度。The single factor test and orthogonal design were used to establish and optimize the ISSR-PCR reaction system of Fraxinus hupehensis Chiú,Shang et Su. The results showed that the appropriate PCR reaction system consisted of 40 ng template DNA,0.7 μmol / L primer,and 7.5 μL 2 ×Taq PCR Plus Master Mix.14 high-efficient polymorphic primers were screened out based on the optimal ISSR reaction system.Furthermore,the ideal annealing temperature of the every selected primer was decided according to the amplified results with different temperatures.

关 键 词：对节白蜡(Fraxinus hupehensis Chiú Shang et Su) ISSR-PCR 反应体系 单因子试验 正交设计 

分 类 号：Q78[生物学—分子生物学] Q943.2