林业科技开发2012，Vol.26Issue(2)：29-32,4.DOI:10.3969/j.issn.1000-8101.2012.02.009
南方红豆杉ISSR -PCR反应体系的建立与优化
An orthogonal design to optimize ISSR-PCR reaction system for Taxus wallichiana var.chinensis
摘要
Abstract
Several important parameters influencing ISSR-PCR amplification on DNA from Taxus wallichiana var. Chinensis were studied with an orthogonal design by four factors and four levels to establish the optimum ISSR reaction system. The optimal ISSR-PCR reaction system was established as a total volume of 20μL consisting of 60ng template DNA, 10 x PCR buffer 2. 0μX, 25 mmol/L MgCl2 l.0μX, 2.5 mmol/L dNTPs l.0μL, 10μmol/L primer 1.0 uX and Taq DNA polymerase 0.2U. The suitable PCR procedure is one cycle denaturing at 94℃ for 4min,then, 41 cycles which involves denaturing at 941 for 30s,annealing at 54.5℃ for 45s and extending at 72℃ for 80s, one cycle extending at 72℃ for 5 min.and kept at4℃o关键词
南方红豆杉/ISSR/体系优化/正交设计Key words
Taxus wallichiana var. Chinensis/ ISSR/ system optimization/ orthogonal design引用本文复制引用
廖盼华,汪庆,李亚,姚淦,杨如同,王淑安..南方红豆杉ISSR -PCR反应体系的建立与优化[J].林业科技开发,2012,26(2):29-32,4.基金项目
中国科学院知识创新工程重要方向项目(编号:KSCX2-YW-Z-0921). （编号:KSCX2-YW-Z-0921）
