观光木SSR-PCR反应体系优化及引物筛选  被引量:4

Optimization of SSR-PCR Reaction System and Prime Screening for Tsoongiodendron odorum

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作  者:黄芳芳[1] 何长青[1] 闫丽君[1] 汪灵丹[1] 徐刚标[1] 

机构地区:[1]中南林业科技大学林学院林木遗传育种实验室,湖南长沙410004

出  处:《中南林业科技大学学报》2015年第10期142-146,共5页Journal of Central South University of Forestry & Technology

基  金:国家林业公益性行业科研专项(201104033)

摘  要:在单因素试验基础上,采用正交试验优化影响观光木SSR-PCR扩增的5种主要影响因素,获得最佳反应体系(10μL)为:Mg2+1.0 mmol,d NTP 0.20 mmol,引物0.25μmol,Taq DNA聚合酶0.75 U,模板DNA 60ng。利用优化SSR-PCR扩增体系,从12对引物中筛选出5对多态性高、重复性好的引物。这为进一步开展观光木种群遗传多样性研究奠定了前期实验基础。In this paper, the main factors affecting SSR-PCR system of Tsoongiodendron odorum were optimized through orthogonal design with the samples of T. odorum, which is based on single factor experiments. SSR-PCR amplytications were performed in a final volume of 10 μL containing 1.0 mmol Mg^2-., 0.20 mmol dNTP, 0.25 μmol of each primers, 0.75 U Taq DNA polymerase, 60 ng of genomie DNA. Using the optimized SSR-PCR system 5 primes with high polymorphism and reproducible properties were selected from 12 primes. This paper establishs the foundation for further study of population genetic diversity in T. odorum.

关 键 词:观光木 SSR 体系优化 引物筛选 

分 类 号:S718.46[农业科学—林学]

 

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