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以改进的SDS法提取褐黄孢链霉菌基因组DNA,对其进行随机扩增多态性(RAPD)研究。通过单因素和正交试验相结合的方法,建立了适于褐黄孢链霉菌RAPD分析的PCR反应体系:包括Taq聚合酶、Mg~(2+)、随机引物、dNTPs和模板浓度。在20μl体系中,模板DNA浓度60~150mg,Taq聚合酶为1.0~1.5U,引物浓度0.3~0.4mmol/L,dNTPs浓度200~250μmol/L,Mg~(2+)浓度2.5~3.0mmol/L。在此反应体系下可扩增出条带数目多且清晰稳定的电泳图谱。
The genomic DNA of Streptomyces taurus was extracted by the improved SDS method, and its was amplified by random amplified polymorphic DNA (RAPD). The PCR reaction system suitable for RAPD analysis of Streptomyces alternaria was established by the combination of single factor and orthogonal test: including Taq polymerase, Mg 2+, random primers, dNTPs and template concentration. The template DNA concentration was 60-150 mg, Taq polymerase 1.0-1.5 U, primer concentration 0.3-0.4 mmol / L, dNTPs concentration 200-250 μmol / L, Mg 2+ concentration 2.5-3.0 mmol / L. In this reaction system can amplify a number of bands and clear and stable electrophoresis patterns.