油茶SRAP-PCR反应体系的建立

以油茶品种大别山2号和赣兴46为试验材料，利用Me5Em8正反向引物组合进行了SRAP-PCR反应体系的L9(34)正交试验，采用正交设计直观分析及方差分析对影响SRAP反应的Taq聚合酶、Mg2+、dNTP和引物浓度进行分析，并对模板DNA浓度进行了单因素试验分析。结果表明，油茶SRAP-PCR 20 μL反应体系的最佳组合为: Taq聚合酶1.20 μmol/min、Mg2+浓度125 mmol/L、dNTP浓度0.15 mmol/L、Primer浓度0.60 μmol/L、模板DNA含量60 ng，并含有2 μL 10×buffer(Mg2+ free)。各因素对油茶SRAP-PCR反应的影响大小依次为:dNTP、 Mg2+ 、Taq聚合酶、Primer。

Establishment reaction system of the SRAP-PCR in Camellia oleifera

In this paper,orthogonal design to SRAP-PCR system for Camellia oleifera cultivar Dabieshan 2 and Ganxing 46 was conducted with the primers combination of Me5 and Em8.Orthogonal design-direct analysis and variance analysis were applied to optimize SRAP-PCR amplification system in 4 factors such as Taq DNA polymerase,Mg2+,dNTP and primer.Template DNA was also analyzed by the single element test.The results indicated that an optimal reaction system(20 μL)of SRAP-PCR system was established: 1.20 μmol/min Taq DNA polymerase 1.25 mmol/L Mg2+,0.15 mmol/L dNTP,0.60 μmol/L primer,60 ng template DNA and 2 μL 10 buffer(Mg2+ free).And,the order of each factor levels affected on the result of SRAP-PCR was dNTP,Mg2+,Taq DNA polymerase and primer.