观光木SSR-PCR反应体系优化及引物筛选

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

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.