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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。
In order to study the genetic background and phylogenetic relationship of Prunella vulgaris germplasm resources, this study used L16 (45) orthogonal design to study the influence of five factors such as Mg 2+ on PCR amplification influences. The results showed that the optimum conditions of SCo T-PCR reaction (20 μL) were 5 Mg 2+, d NTPs, Taq DNA polymerase, primers and template DNA The concentrations were 2.188 mmol / L, 0.113 mmol / L, 1.0 U, 0.625 μmol / L and 30 ng, respectively. Based on this, 12 SCo T primers with stable amplification and clear bands were initially screened from 18 primers. The SCo T-PCR reaction system was established. The results of 18 primers and 13 germplasm resources showed that the system has good stability and high reliability, and can meet the research on the molecular level of petals.