晒烟RAPD反应体系的优化

Optimization of RAPD reaction system in sun-cured tobacco

以晒烟苗期幼叶为试材,通过设置不同的模版DNA、引物、dNTP、Mg2+及Taq DNA聚合酶的浓度梯度,优化RAPD的反应条件,建立1个适合晒烟的比较稳定的RAPD反应体系。结果表明,适合晒烟RAPD的反应体系是在25μL反应体系中,含模板40ng;引物UBC-388 1.5μL;Mg2+1.6mM;dNTP 0.2mM;Taq酶1U。

To establish a more stable RAPD reaction system for sun-cured tobacco,the young leaves of sun-cured tobacco was used to study optimization of RAPD reaction conditions through setting different concentration gradient of template DNA,primers,dNTP,Mg^2＋,and Taq DNA polymerase（hereinafter referred to as Taq polymerase）.The results showed that the suitable RAPD reaction system for sun-cured tobacco is that in 25 μL reaction system containing 40 ng template;1.5 μL primer UBC-388;1.6 mM Mg^2＋,0.2 mM dNTP and 1 U Taq polymerase.