摘要
为研究距瓣尾囊草种质资源遗传背景和系统进化提供理论依据和技术支持,本研究利用L16(45)正交设计,研究Mg^(2+)等5个因素对PCR扩增结果的影响。研究结果表明,在距瓣尾囊草SCo T-PCR的最优反应体系(20μL)中,Mg^(2+)、d NTPs、Taq DNA聚合酶、引物以及模板DNA等5个因素的最优浓度分别是2.188 mmol/L、0.113 mmol/L、1.0 U、0.625μmol/L和30 ng。在此基础之上,从18条引物中初步筛选出12条扩增结果稳定、条带清晰的SCo T引物。建立了距瓣尾囊草SCo T-PCR反应体系,经过18条引物和13份种质资源的验证,证明体系稳定性好、可靠性高,能满足距瓣尾囊草分子水平上的研究。
Orthogonal design of L_(16)(4~5) was applied to optimize SCo T-PCR amplification system of Urophysa rockii Ulbr in five factors such as Mg2+, d NTPs, primer, Taq DNA polymerase and template DNA. An optimal reaction system of Urophysa rockii Ulbr was established. PCR reaction mixtures contained Mg2+2.188 mmol/L, d NTPs0.113 mmol/L, Taq DNA polymerase 1.0 U, primer 0.625 μmol/L and template DNA 30 ng. 18 SCo T primers and13 different Urophysa rockii Ulbr populations were used to test the optimized reaction system. Ideal results with high stability and clear polymorphic bands were obtained. The SCo T system established in this reportis useful for genetic diversity analysis of Urophysa rockii Ulbr, which possesses considerable potential for Urophysa rockii Ulbr breeding.
出处
《分子植物育种》
CAS
CSCD
北大核心
2016年第9期2453-2459,共7页
Molecular Plant Breeding
基金
国家自然科学基金(31400333)
西南科技大学实验室开放基金(15xnkf02)
西南科技大学大学生创新基金(CX16-066)
重庆市科委重大项目(cstc2013yykfc80002)共同资助
关键词
距瓣尾囊草
SCoT标记
正交设计
遗传背景
Urophysa rockii Ulbr
Start codon targeted polymorphism
Orthogonal design
Genetic analysis
