安徽农业科学2009，Vol.37Issue(31)：15163-15166,4.
珍稀濒危植物大果木莲ISSR—PCR反应体系的建立
Establishment of ISSR-PCR Reaction System for the Rare and Endangered Plant Manglietia grandis Hu et Cheng
摘要
Abstract
[Objective] The study aimed to establish the effective and stable ISSR-PCR reaction system for Manglietia grandis Hu et Cheng. [Method] With the leaves of rare and endangered plant M. grandis as tested material, based on high quality DNA extracted by high salt precipitation method, the single factor grade experiment with 5 factors in ISSR-PCR reaction system was taken to optimize and finally establish the effective and stable ISSR-PCR reaction system through single-factor gradient test. [ Result] The effective and stable ISSR-PCR reaction system for M. grandis was established,which contained 10 × buffer 2.0 Ul,Mg~(2+) 2.0 mmol/L,Taq enzyme 0.5 U.DNA template 40 ng,primer 0.8 μmol/L,dNTPs 0.2 mmol/L,ddH_2O 13. 3μl in total 20.0 μl system. The PCR amplification program was that the initial denaturation was for 5 min at 94℃ .followed by 40 cycles of denaturing at 94℃ for 1 min,annealing at 50 -60℃ for 45 s,extension at 72℃ for 2 min,and a final extension was at 72 ℃. for 7 min. [ Conclusion ] The study provided the theoretical basis for further opening out the genetic diversity and structure and making out the scientific protection measures.关键词
珍稀濒危植物/大果木莲/基因组DNA/ISSR-PCR反应体系Key words
Rare and endangered plants/ Manglietia grandis Hu et Cheng/ Genomic DNA/ ISSR-PCR reaction system分类
农业科技引用本文复制引用
毛云玲,陈少瑜,韩燕,吴涛,司马永康,郝佳波,付玉嫔..珍稀濒危植物大果木莲ISSR—PCR反应体系的建立[J].安徽农业科学,2009,37(31):15163-15166,4.基金项目
云南省自然科学基金项目(2006C0066M). （2006C0066M）
