期刊论文

中文

经济林研究

1003-8981

2016

34

1

33-39,44

10.14067/j.cnki.1003-8981.2016.01.006

(201304807):国家林业公益性行业科技专项项目“仙茅、鸡血藤资源培育及林下种植技术研究”

本校为第一机构

为了改进仙茅基因组DNA的提取方法,从而为其遗传多样性研究提供参考依据,以仙茅根茎为材料,采用CTAB法探讨了仙茅根茎DNA大量提取中CTAB浓度、PVP用量及Na Cl浓度对其DNA质量的影响情况。结果表明,仙茅根茎DNA的大量提取方法的最优组合为:CTAB的浓度为2%,PVP的加入量为0.20 g,Na Cl的浓度为4 mol/L。为了得到仙茅的最佳ISSR-PCR反应体系,以（CT）8 RG为引物,采用单因素实验与正交实验相结合的方法,得到的最优反应体系（25μL）为:1×PCR Buffer,模板DNA浓度为50 ng,Mg2+浓度为2.50 mmol/L,d NTPs浓度为0.20 mmol/L,引物浓度为0.40μmol/L,Taq DNA聚合酶浓度为1.50 U。

In order to improve extraction method of genomic DNA in Curculigo orchioides, and to provide some references for researches on genetic diversity of C. orchioides, taking tubers in C. orchioides as materials, effects of CTAB concentration, PVP dosage and Na Cl concentration on quality of extracted DNA by using CTAB method. The results showed that the optimal DNA extraction combination of tuber in C. orchioides was 2% CTAB, 0.20 g PVP and 4 mol/L Na Cl. To gain the optimal ISSR-PCR reaction system, taking（CT）8RG as primer, some single factor ex...