麻疯树cpSSR标记技术的建立与体系优化研究

摘要

[Objective] The aim was to build an optimal cpSSR-PCR system for Jatropha curcas Linn. [Method] Optimize cpSSR-PCR amplification system for Jatropha curcas Linn in terms of five factors( Taq DNA polymerase, Mg~(2+), DNA template, dNTP and primer)from several levels. [Result] An optimal cpSSR-PCR system for Jatropha curcas Linn was obtained as 2.00 mmol/L Mg~(2+), 2 U/μl Taq DNApolymerase, 0.2 mmol/L dNTP, 0.2 μmol/L Primer, 35 ng/μl DNA template for 20 μl reaction syetem. [Conclusion] The optimal annealing temperature for cpSSR-PCR reaction syetem is determined as 52 ℃. The cpSSR reaction system was steady and reproducible.%[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(Taq DNA聚合酶、DNA模板、Mg~(2+)、dNTP、引物)进行优化试验,筛选各反应因素的最佳水平.[结果]20 μl反应体系各组分的最合适浓度分别为10×Buffer、2.00 mmol/L Mg~(2+)、2 U/μl Taq DNA聚合酶、0.2 mmol/L dNTP、0.2 μmol/L引物、35 ng/μl DNA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.