安徽农业科学2009，Vol.37Issue(22)：10424-10425,10428,3.
大叶栎ISSR扩增体系的优化
Optimization of ISSR Amplification System of Castanopsis fissa
摘要
Abstract
[Objective] The study aimed to discuss factors that affected ISSR amplification effect of Castanopsis fissa and establish optimized system of ISSR amplification for C. fissa. [Method] With tender leaves of C. fissa as tested materials, genomic DNA of C. fissa was extracted with modified CTAB method and the DNA conc. was diluted to 30 ng/μl. Single-factor experiment was designed to test the effect of the dosages of DNA template, PCR buffer and Taq DNA polymerase, as well as the final conc. of substrate dNTP and primers on ISSR amplification. [Result] The optimized system of ISSR amplification for C. fissa was as follows: the total 20 μl reaction volume contained DNA template of 60 ng, PCR buffer 10×buffer of 2.0 μl, the final substrate dNTP conc. was 0.25 μmol/L, the final primer conc. was 0.3 μmol/L and Taq polymerase dosage was 1 U. In the system, the clear bands with high polymorphism could be amplified, which could meet the requirements of ISSR diversity analysis for C. fissa. [Conclusion] The optimized system of ISSR amplification for C. fissa was established and provided the technical foundation for studying the population genetic diversity of C. fissa population in Guangxi Province.关键词
大叶栎/ISSR/反应体系/优化Key words
Castanopsis fissa/ISSR/Reaction system/Optimization分类
农业科技引用本文复制引用
梁文汇,谭健晖,黄寿先,梁机,谢素平,刘光金,黄婷,周传明..大叶栎ISSR扩增体系的优化[J].安徽农业科学,2009,37(22):10424-10425,10428,3.基金项目
广西自然科学基金(桂科字0542024) （桂科字0542024）
广西"十五"林业科技项目(林科字2002[18]). （林科字2002[18]）
