论文部分内容阅读
为了加强对野生山茴香物种资源的保护,本研究在单因素试验基础上,对山茴香SRAP-PCR反应体系进行了正交设计优化,对主要影响因素:Taq酶、d NTPs、Mg~(2+)、引物进行了4因素3水平的正交设计试验L_9(3~4)。研究表明,最佳反应体系为:Taq聚合酶0.06 U/μL,d NTPs 0.6 mmol/L,Mg~(2+) 1.5 mmol/L,引物0.1μmol/L,总体积25μL。利用最优体系进行了引物筛选,从30对SRAP引物中获得了15对条带清晰,多态性良好的引物组合。本研究为利用SRAP技术进行山茴香种质资源分析、分子遗传育种等研究奠定了技术基础。
In order to strengthen the protection of wild mountain fennel species resources, this study, based on single factor experiments, optimized the SRAP-PCR reaction system of mountain fennel by orthogonal design. The main influencing factors were Taq enzyme, dNTPs, Mg ~ (2) +). Primers were subjected to 4 factors and 3 levels of orthogonal design experiment L_9 (3 ~ 4). The results showed that the optimal reaction system was Taq polymerase 0.06 U / μL, d NTPs 0.6 mmol / L, Mg 2+ 1.5 mmol / L, Primer 0.1 μmol / L and total volume 25 μL. Fifteen pairs of SRAP primers were screened by the optimal system. Fifteen pairs of primers with clear bands and good polymorphism were obtained. This study lays the technical foundation for the utilization of SRAP technology for the analysis of fennel germplasm resources and molecular genetic breeding.