黄曲霉菌ISSR反应体系的建立与优化

Establishment and optimization of ISSR-PCR reaction system for Aspergillus flavus

为建立和优化黄曲霉菌ISSR反应体系,在单因素实验的基础上,运用正交试验确定了黄曲霉ISSR-PCR的最优体系为10×PCR buffer 2.0μL、1.4 U Taq DNA聚合酶、0.35 mmol/L dNTP、0.7μmol/L引物、25 ng模板DNA、1.75 mmol/L MgCl2。该体系的建立为利用ISSR技术对黄曲霉菌进行遗传多样性分析、遗传图谱构建奠定了技术基础。

Effect for ISSR-PCR reaction of Aspergillus flavus on five factors （Taq DNA polymerase, dNTP, primer, genomic DNA, Mg2＋） were analysed by orthogonal design for establishing and optimizing ISSR-PCR reaction system of A. fiavus. The results showed the optimal reaction system of ISSR- PCR amplification （20μL） for A. flavus was composed of 2.0μL of 10×reaction buffer, 1.4 UTaq DNA polymerase, 0.35 mmol/L dNTPs, 0.7 pmol/L primer and 25 ng genomic DNA, 1.75 mmol/L MgCl2.