【摘要】 对麻栎基因组DNA的SRAP-PCR体系中dNTP、Taq酶、Mg2+、引物、模板DNA进行正交试验优化,结果表明最佳反应体系为dNTP浓度为0.3 mmol.L-1,Taq酶1.5U,Mg2+浓度为2mmol.L-1,引物浓度0.2μmol.L-1,模板DNA40ng(20μL反应体系)。运用优化体系,从110对SRAP引物组合中,筛选出多态性较好的8对SRAP引物,对8个不同地区麻栎进行SRAP标记分析。共检测到45个多态性位点,多态性条带百分比为51.72%。应用NTSYS-pc软件进行聚类分析(UPGMA),建立了麻栎亲缘关系树状图,表明SRAP可有效用于麻栎种质资源鉴定与遗传多样性分析。更多还原

【Abstract】 The concentration of dNTP,Taq DNA polymerase,Mg2+,primer and DNA template in the SRAP-PCR system was optimized for genomic DNA in Quercus acutissima Carr.The optimized concentrations were dNTP at 0.3 mmol·L-1,Taq DNA polymerase at 1.5U,Mg2+ at 2mmol·L-1,primer at 0.2μmol·L-1,and DNA at 40ng,with a total volume of 20 μL.Eight optimized polymorphic SRAP primer combinations were obtained with the optimized system from 110 SRAP primer combinations.The molecular identification of 8 different regions Quercus acutissima Carr was conducted with the SRAP marker system.55 polymorphic loci was detected,the percent of polymorphic band was 51.72%.The relative relationship tree of Quercus acutissima Carr was constituted by Cluster analysis(UPGMA).The results indicated that SRAP would play an important role in Quercus acutissima Carr germ plasma identification and genetic diversity analysis.更多还原