漾濞泡核桃 ISSR-PCR 反应体系的建立和优化

摘要

以漾濞泡核桃优株为研究对象，研究其ISSR-PCR反应体系中的5个影响因子（模板DNA浓度、Taq DNA 聚合酶浓度、引物浓度、dNTPs浓度、Mg2＋浓度）对PCR扩增效果的影响，从而建立起漾濞泡核桃的最佳ISSR-PCR反应体系。结果表明：模板DNA最佳浓度为50 ng/（25μL），Taq聚合酶浓度为0.75U/（25μL），引物浓度为0.7mmol/L，dNTPs浓度为0.20mmol/L，Mg2＋浓度为2.0mmol/L，利用该最佳体系对试验中所选取的样本进行扩增反应。建立了适宜于漾濞泡核桃的ISSR-PCR扩增的最佳体系，该体系的建立为利用ISSR分子标记技术对漾濞泡核桃进行种质资源研究奠定了基础。%The effects of template DNA concentrations , Taq DNA polymerase concentrations , primers concentrations , dNTPs and Mg2+concentrations in ISSR-PCR reaction system on PCR amplification have been studied , and optimum ISSR-PCR reaction system was set up.Results showed that the most suitable concentrations of template DNA, Taq DNA polymerase, primers, dNTPs and Mg2+ were 50ng/(25μL), 0.75U/(25μL), 0.7mmol/L, 0.20mmol/L and 2.0mmol/L respectively.The optimal IS-SR-PCR reaction system for Juglans sigillata established in this research provides a basis for using ISSR molecular marker technology to study the germplasm resources of Juglans sigillata.