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[目的]建立麻疯树cpSSR-PCR反应的最佳反应体系。[方法]对影响麻疯树cpSSR-PCR反应体系的5个因素(TaqDNA聚合酶、DNA模板、Mg2+、dNTP、引物)进行优化试验,筛选各反应因素的最佳水平。[结果]20μl反应体系各组分的最合适浓度分别为10×Buffer、2.00mmol/LMg2+、2U/μlTaqDNA聚合酶、0.2mmol/LdNTP、0.2μmol/L引物、35ng/μlDNA模板。[结论]最佳的退火温度为52℃;该反应体系的稳定性和可重复性均较好。
[Objective] The research aimed to establish the optimal reaction system of cpSSR-PCR for Jatropha curcas. [Method] Five factors (Taq DNA polymerase, DNA template, Mg2 +, dNTP and primer) that affected the cpSSR-PCR reaction system of Jatropha curcas were optimized and the optimum level of each factor was screened. [Result] The optimal concentration of each component in 20μl reaction system was 10 × Buffer, 2.00mmol / LMg2 +, 2U / μl Taq DNA polymerase, 0.2mmol / LdNTP, 0.2μmol / L primer and 35ng / μl DNA template, respectively. [Conclusion] The best annealing temperature was 52 ℃. The stability and repeatability of the reaction system were good.