基于琼脂糖凝胶电泳的小麦SSR扩增体系优化

Optimization of SSR Amplification System and Agarose Gel Electrophoresis of Amplificons

以小麦基因组DNA为试验材料,探索SSR扩增反应中5种反应组分(模板DNA、dNTPs、引物、Mg2+和TaqDNA聚合酶)对SSR扩增结果的影响。研究发现,引物、Mg2+和TaqDNA聚合酶对PCR扩增结果的影响最显著,其次是模板DNA,dNTP对PCR扩增结果的影响不明显。此外,借助该优化的SSR反应体系,对小麦7B染色体上的多对SSR引物进行了多态性筛选。试验结果表明,优化的SSR扩增体系结合高浓度琼脂糖凝胶能够对显性或差异显著的共显性标记进行高效分离。

In this study,effect of five reaction components,including template DNA,dNTPs,primer,Mg2＋ and Taq DNA polymerase,on the SSR amplification results were studied using wheat genomic DNA as PCR template.Mg2＋ and Taq DNA polymerase showing most significant influence on PCR products,and then is template DNA,dNTPs have no significant effect on it.Furthermore,SSR markers on wheat chromosome 7B were screened by the optimized SSR system.Results indicated that,dominant SSR markers or codominant markers with significant difference on length can be separated by optimized SSR amplification system and high concentration agarose gel effectively.