安徽农业科学2009，Vol.37Issue(21)：9869-9871,3.
青海云杉ISSR-PCR反应体系的建立与优化
Establishment and Optimization of ISSR-PCR Reaction System for Picea crassifolia kom.
王文峰 1李毅 1马彦军 1李栋栋1
作者信息
- 1. 甘肃农业大学林学院,甘肃兰州,730070
- 折叠
摘要
Abstract
[Objective]The experiment aimed to determine the optimum ISSR-PCR reaction system of Picea crassifolia kom.. [Method]Picea crassifolia kom. was used as material to select and optimize influencing factors of ISSR-PCR such as Mg2+, dNTPs, TaqDNA polymerase, template DNA, primers, annealing temperature. [Result]The optimum ISSR-PCR reaction system in 20 μl reaction system was consisted of 1 μl 10×buffer, 1.5 mmol/L Mg2+, 0.2 mmol/L dNTPs, 1.0 U TaqDNA polymerase, 40 ng template DNA, 0.6 μmol/L primers. According to gradient test of annealing temperature in optimum ISSR-PCR reaction system of Picea crassifolia kom., it was found that the optimum annealing temperature of UBC818 was 54.2 ℃ and the annealing temperature was different for different primers.[Conclusion]The construction of ISSR-PCR reaction system provided technical basis for classification of germplasm resources, construction of genetic map, gene mapping of Picea crassifolia kom. through using ISSR technology.关键词
青海云杉/ISSR/体系建立/优化Key words
Picea crassifolia kom./ISSR/System establishment/Optimization分类
农业科技引用本文复制引用
王文峰,李毅,马彦军,李栋栋..青海云杉ISSR-PCR反应体系的建立与优化[J].安徽农业科学,2009,37(21):9869-9871,3.
