摘要
为建立并优化适用于芒草的ISSR-PCR扩增反应体系,进一步研究野生芒草群体的遗传多样性水平。以吉林省采集的芒草(Miscanthus sinensis)为材料,采用单因子试验的方法研究模板DNA、Taq DNA聚合酶的用量及引物浓度和退火温度对PCR扩增的影响。结果显示:在20μL反应体系中,含有模板DNA 40 ng,d NTPs 0.4 mmol/L,引物0.6μmol/L,Taq DNA聚合酶1.5 U。此外,筛选到10条扩增稳定、条带丰富的候选引物,并确定了各自的最佳退火温度。
The aims were to establish and optimize the amplification system of ISSR-PCR and further study the genetic diversity of wild Miscanthus sinensis populations. M. sinensis samples were collected from Jilin Province for the test. The influence of DNA template, Taq DNA polymerase, primer and annealing temperature on the PCR amplification was studied by the single factor test method. The results indicated that： 20 μL reaction system contained 40 ng template DNA, 0.4 mmol/L dNTPs, 0.6 μ mol/L primer, Taq DNA polymerase 1.5 U. 10 ISSR primers with stable amplification bands and rich polymorphism for ISSR-PCR were selected, and the optimal annealing temperature was also found.
出处
《中国农学通报》
2015年第33期186-191,共6页
Chinese Agricultural Science Bulletin
基金
吉林省教育厅"十二五"科学技术研究项目"紫花苜蓿细胞质雄性不育系与保持系SCAR标记的鉴定研究"(吉教科合字[2015]第197号)
