【摘要】 以红三叶(Trifolium pretense L.)、白三叶(Trifolium repens L.)和杂三叶(TrifoliumHybridumL.)三种三叶草为材料,对RAPD反应条件中的dNTP浓度、随机引物浓度、模板DNA用量、Taq DNA聚合酶用量、退火温度以及预变性时间分别进行梯度试验。结果表明,该反应体系体积为25μL:ddH2O,19.1μL;10×Buffer,2.5μL;Primer,200 nM;dNTP,200μM;Taq DNA聚合酶,1U;Template DNA 1ng/μL。通过试验建立了一套稳定的RAPD反应体系,该反应体系扩增效果良好。更多还原

【Abstract】 Three species of clover were used as the materials of extracting DNA in this experiment,they were Trifolium pretense L.、Trifolium repens L. and Trifolium Hybridum L..The experiment step to the concentration of dNTP、random primers、template DNA、Taq DNA polymerase、annealing temperature and preheat denaturalization time on RAPD were conducted.Then a stable reaction system of RAPD was set up.In the optimalization of RAPD system with totale volume of 25μL: ddH₂O,19.1μL;10×Buffer,2.5μL;Primer,200nM;dNTP,200μM;Taq DNA E,1U;Template DNA 1ng/μL.The effect of amplifying with the reaction system was the most optimal.更多还原