摘要
对广西红锥种群的DNA进行提取和ISSRPCR扩增体系进行优化。分析了退火温度、模板DNA浓度、Mg2+浓度、dNTPs浓度、TaqDNA聚合酶用量对反应结果的影响。红锥ISSRPCR分析较适宜的扩增体系是:25μL PCR反应体积中,buffer(10 mM TrisHCl,pH 9.0,50 mM KCl,0.1%Triton X100),1.25 U Taq DNA聚合酶,4种dNTPs各0.2 mM,0.4μM引物,2.0mM MgCl2,50 ng模板DNA。
The DNA and ISSR-PCR amplification of Guangxi Catamopsis hystrix was extracted and optimized respectively. The factors which affected the ISSR amplification such as annealing temperature, template DNA dosage, Mg^2＋ concentration, dNTPs concentration and unit of Taq DNA polymerase were selected and optimized. The results showed that the reaction system being suitable for ISSR - PCR of Castanopsis hystrix was as follows ： 1 × Taq buffer （10 raM/L Tris-HCl , pH 9.0, 50 mM/L KC1 and 0.1% Triton X-100） , 1.25 U Taq DNA polymerase , 0.2 mM 4 8 dNTP, 0.4μM primers, 2.0 mM/L MgCl2 and 50 ng template DNA in total 25 μL reaction volume.
出处
《广西林业科学》
2011年第4期288-291,共4页
Guangxi Forestry Science
基金
广西林科院基本科研业务费项目(林科200901)
关键词
红锥
ISSR
优化
遗传多样性
Castanopsis hystrix
ISSR
optimization
genetic diversity
