植物研究2009，Vol.29Issue(1)：80-85,6.
利用正交设计优化兴安落叶松RAPD-PCR反应体系
Establishment and Optimization of RAPD-PCR Reaction System for Larix gmelnii( Rupr. ) Rupr Using Orthogonal Design
摘要
Abstract
The orthogonal design was used to optimize RAPD amplification system of Larix gmelnii ( Rupr. ) Ru-pr with five factors ( Taq polymerase, Mg~(2+) , dNTP, primer, DNA templet) at four levels, respectively. Through the deep analysis, a suitable RAPD-PCR reaction system was established, namely 20 μL reaction system contai-ning 1.0 U Taq DNA polymerase, 2.5 mmol·L~(-1) Mg~(2+) ,0.25 mmol·L~(-1) dNTP, 0.5 μmol·L~(-1) primer, 1× PCR buffer, 90 ng DNA template. 20 primers with stable amplification and rich polymorphism for RAPD were screened. The optimal annealing temperature for RAPD-PCR reaction was proposed by gradient PCR. The result provided a standardizing RAPD-PCR program for the analysis of genetic diversity of L.gmelnii(Rupr. ) Rupr.关键词
兴安落叶松/RAPD-PCR/正交设计/反应体系Key words
Larix gmelnii(Rupr. )Rupr/RAPD-PCR/orthogonal design/reaction system分类
农业科技引用本文复制引用
李雪峰,张含国,贯春雨,张瑶..利用正交设计优化兴安落叶松RAPD-PCR反应体系[J].植物研究,2009,29(1):80-85,6.基金项目
国家科技基础性工作专项(2007FY110400-3) （2007FY110400-3）
国家科技支撑项目(2006BAD01A1401-01) （2006BAD01A1401-01）
国家自然科学基金项目(30640034) （30640034）
