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本研究以催吐萝芙木为材料,利用正交试验设计对影响SRAP-PCR反应的不同浓度模板DNA、Taq DNA聚合酶、引物和d NTP进行优化,建立催吐萝芙木的SRAP-PCR最佳反应体系。20μL反应体系包括:10×Taq Plus Buffer(含1.5 mmol/L Mg2+)2μL,d NTP 0.2 mmo1/L,Taq DNA聚合酶0.75 U,模板DNA 40 ng、正/反引物各1.2μmo1/L。在此条件下,对能够在催吐萝芙木、四叶萝芙、蛇根木、苏门答腊萝芙木、云南萝芙木中扩增的引物进行了筛选,获得30对具有通用性扩增的引物,为后续通过SRAP探讨其遗传多样性和亲缘关系方面的研究提供参考。
In this study, we used the orthogonal design to optimize the SRAP-PCR of different concentrations of template DNA, Taq DNA polymerase, primers and d NTP affecting the SRAP-PCR reaction. reaction system. 20μL reaction system consisted of 2μL of 10 × Taq Plus Buffer (containing 1.5 mmol / L Mg2 +), 0.2 mmol / L of dNTP, 0.75 U of Taq DNA polymerase, 40 ng of template DNA and 1.2 μmol / L of each primer. Under these conditions, the primers that could amplify in Rattus rotundus, Spartina alterniflora, Snake rootstock, Sumatra radish wood, and Yunnan Rauwolf wood were screened, and 30 pairs of primers with universal amplification were obtained, Which will provide reference for further research on its genetic diversity and kinship through SRAP.