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.
出处
《中南林业科技大学学报》
CAS
CSCD
北大核心
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)
关键词
观光木
SSR
体系优化
引物筛选
Tsoongiodendron odorum
SSR
Reaction system optimization
Prime screening