正交设计优化烟草SSR反应体系

The Optimization of SSR Analysis System in Tobacco by Orthogonal Design

以烤烟品种K326为材料,探讨影响烟草SSR扩增的各种因素,建立能够稳定扩增烟草基因组的体系,为研究烟草重要性状的遗传基础及建立分子辅助标记育种的技术平台奠定基础。结果表明:在总体积为20μl的PCR反应中,含35 ng模板DNA,1.5U聚合酶,MgCl2终浓度为1.5 mmol/L,dNTPs浓度为200μmol/L,引物浓度为0.05μmol/L时效果较好。

Using flue-cured tobacco Vc K326 as material, this paper studied the factors influencing the amplification of SSR system in tobacco and established a stable SSR reaction system to lay a foundation for molecular marker assistant breeding of tobacco and the effects of major elements in PCR reaction system on SSR analysis were studied. The results showed that under the conditions of 1.5U TaqDNA polymerase,terminal MgCl2 concentration of 1.5 mmol/L,350 μmol/L dNTPs, 0.05 μmol/L primers and 35ng templates of DNA in total volume of 20 ul, the result was better in PCR reaction.