利用正交设计L16(45)对萝卜SSR - PCR反应体系的5因素(Taq酶、Mg2+、模板DNA、dNTP、引物)在4个水平上进行优化,获得了最佳反应体系,即20μl反应体系中含有0.10 mmol/L dNTP,3.0 mmol/LMg2+,50 ng模板,0.500 mol/L引物,1.0 U Taq酶.使用优化的反应体系,扩增条带清晰,可满足不同引物组合和萝卜材料的要求.此研究为今后利用SSR技术对萝卜种质资源进行分类、构建遗传图谱和基因定位奠定了基础.%The orthogonal design L16(45)was used to optimize the EST - SSR system for radish at four levels of five factors (Dntp, primer, Mg2 + , Taq DNA polymerase and template DNA). The 20 (xl optimal SSR -PCR reaction system included 0. 10 mmol/L Dntp,3. 0 mmol/L Mg2+ , 50 ng template DNA,0. 500 fimol/L primer and LOU Taq DNA polymerase. With the optimal system, the obtained bands were clear and could meet the requirements of different primer combinations and radish materials. This study provided basis for the analysis of diversity, genetic map construction and gene localization of radish germplasm resources by SSR technique.
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