广西特有植物小花异裂菊ISSR-PCR反应体系的建立

摘要

以小花异裂菊的叶片为材料,采用正交和单因素试验研究Mg2+、dNTP、TaqDNA聚合酶、引物、模板DNA浓度对ISSR-PCR扩增效果的影响.建立了适宜于小花异裂菊的ISSR-PCR反应体系,即25μL的体系中含Mg2+ 1.5 mmol/L,dNTP 0.20 mmol/L,TaqDNA聚合酶1.5U,引物0.8μmol/L,模板DNA 50 ng,10×Buffer 2.5μL.该体系稳定、可靠,为利用ISSR分析小花异裂菊遗传多样性等工作奠定了良好的基础.%Through orthogonal design and single factor test, to establish ISSR-PCR program for Heteroplexis sericophylla. Five factors were tested by orthogonal design and single factor test, including Mg2+ concentration, dNTP concentration, Taq DNA polymerase concentration, primer concentration and template DNA concentration. ISSR-PCR reaction system of Heteroplexis sericophylla contained 1. 5 mmol/L Mg + ,0. 20 mmol/L dNTP, 1.5 U Taq DNA polymerase,0. 8 μmol/L primer,50 ng template DNA. The optimized reaction system was suitable for the study of genetic diversity.