论文部分内容阅读
以杜鹃花为试验材料,采用正交实验方法,研究模板DNA浓度、ISSR引物浓度、dNTPs浓度、Taq DNA聚合酶浓度、Mg2+浓度等5个因素对ISSR-PCR反应体系的影响,以建立适合杜鹃花的ISSR-PCR最佳扩增体系。结果表明:杜鹃花ISSR反应体系的最佳条件为模板DNA用量为60ng/20μL,ISSR引物浓度0.60μmol/L,dNTPs浓度0.50μmol/L,Taq DNA聚合酶浓度30U/mL,Mg2+浓度为0.6mmol/L。采用该反应体系可以从10份杜鹃花(R.simsii)基因组内扩增出稳定性高、重复性好ISSR-PCR产物。该研究为杜鹃花的遗传多样性分析、ISSR指纹图谱构建、亲缘关系鉴定等研究奠定了基础。
The effects of template DNA concentration, ISSR primer concentration, dNTPs concentration, Taq DNA polymerase concentration and Mg2 + concentration on ISSR-PCR reaction system were studied by orthogonal test. Flower ISSR-PCR optimal amplification system. The results showed that the optimum conditions of ISSR reaction system were 60ng / 20μL template DNA, ISSR primer 0.60μmol / L, dNTPs 0.50μmol / L, Taq DNA polymerase 30U / mL and Mg2 + 0.6mmol / L. Using this reaction system, ISSR-PCR products with high stability and repeatability can be amplified from 10 genomes of R.simsii. This study laid the foundation for the research on the genetic diversity of rhododendron, ISSR fingerprinting and the identification of kinship.