观光木SSR-PCR反应体系优化和引物筛选.PDF

第35 卷 第10 期 中 南 林 业 科 技 大 学 学 报 Vol. 35 No. 10 2015 年 10 月 Journal of Central South University of Forestry Technology Oct. 2015 Doi:10.14067/j .cnki.1673-923x.2015.10.025 http : // 观光木SSR-PCR反应体系优化及引物筛选 黄芳芳，何长青，闫丽君，汪灵丹，徐刚标 （中南林业科技大学 林学院 林木遗传育种实验室，湖南 长沙 4 10004 ） 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 μ 2+ μ Taq 应体系 （10 L ）为：Mg 1.0 mmol，dNTP 0.20 mmol，引物0.25 mol， DNA 聚合酶0.75 U ，模板DNA 60 ng 。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。 关键词：观光木；SSR ；体系优化；引物筛选 中图分类号：S718.46 文献标志码：A 文章编号：1673-923X(2015) 10-0142-05 Optimization of SSR-PCR Reaction System and Prime Screening for Tsoongiodendron odorum HUANG Fang-fang , HE Chang-qing, YAN Li-jun, WANG Líng-dan, XU Gang-biao (College of Forestry, Central South University of Forestry and Technology, Changsha 410004, Hunan, China) Abstract: 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 Mg2+, 0.20 mmol dNTP, 0.25 μmol of each primers, 0.75 U Taq DNA polymerase, 60 ng of genomic 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. Key words: Tsoongiodendron odorum; SSR; Reaction system optimization; Prime screening 观光木Tsoongiodendron odorum Chun ，属木兰