芝麻SRAP反应体系的建立与优化

Establishment and Optimization of SRAP Reaction System in Sesame

通过对芝麻SRAP反应体系主要影响因素进行优化,建立最佳反应体系。以芝麻幼叶提取的DNA为实验材料,对影响SRAP扩增结果的重要反应因素dNTPs、Mg2+、Taq酶、随机引物及模板DNA设置不同梯度实验,得到了芝麻扩增多态性高、稳定性强、带型清晰的SRAP最佳反应体系为:dNTPs(10mmol/L)0.30μl,Mg2+(25mmol/L)1.20μl,Taq酶1.00U,正反引物各50ng,DNA模板80ng,10×Buffer1.5μl,总体积15μl,为SRAP标记技术在芝麻分子生物学研究方面的应用奠定了基础。

In order to optimize the key factors of SRAP reaction system in sesame （Sesamum indicum L.）, establish an optimum SRAP system, DNA was extracted from young leaf and taken as tested materials, the SRAP amplification system was optimized from many factors in different grads, such as DNA template, Mg2＋, dNTPs, Taq DNA polymerase and primers. The optimum SRAP system （total volume of 15μl） established was as follows： dNTPs （10mmol/L） 0.30μl, Mg2＋ （25mmol/L） 1.20μl, Taq polymerase 1.00U, forward primer 50ng, reverse primer 50ng, DNA template 80 ng and 10×Buffer 1.5μl. The optimum SRAP reaction system could amplify high levels of polymorphism, good repeatability and clear band pattern. This work is a foundation for applying SRAP marker to sesame molecular biology studies.