【摘要】 目的建立稳定性高、重复性好的适合玉竹ISSR-PCR扩增的反应体系。方法以玉竹根茎为材料,采用改良的CTAB法提取玉竹基因组DNA,并利用单因素多水平试验,分别对模板DNA、引物、Mg2+、Taq酶、dNTP的浓度和退火温度进行考察。结果玉竹的最佳反应体系为:在20μL的反应体系中,含1×buffer、15 ng模板DNA、0.4μmol/L引物、2 mmol/L Mg2+、1.0 U Taq酶、200μmol/L dNTP。此反应体系以52℃的退火温度扩增出来的玉竹电泳图谱清晰,重复性好。结论本实验反应体系适宜于玉竹ISSR-PCR的扩增,为玉竹下一步分子生物学的研究打下基础。更多还原

【Abstract】 Objective To establish a high stabilizing reaction system for Polygonatum odoratum ISSR-PCR.Methods The Polygonatum odoratum DNA was extracted with improved CTAB technique and the template DNA concentration,primer concentration,Mg2+ concentration,dNTP concentration,Taq polymerase dosage and temperature were researched respectively.Result The best reaction system of Polygonatum odoratum was in the 20 μL system containing 1×buffer,15 ng template DNA,0.4 μmol/L primer,2 mmol/L Mg2+,1.0 U Taq polymerase,200 μmol/L dNTP.Conclusion This reaction system is suitable to amplification of ISSR-PCR of Polygonatum odoratum and has given good foundation for further study of Polygonatum odoratum.更多还原