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、Mg2+、随机引物、dNTPs和模板浓度。在20μL体系中，模板DNA浓度60-150ng，Taq聚合酶为1.0-1.5U，引物浓度0.3-0.4mmol/L，dNTPs浓度200-250μmol/L，Mg2 +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。
An improved SDS method was used to extract genomic DNA from Streptomyces gilvosporeus. The RAPD conditions were optimized by single factor experiment and orthogonal experiment, including Taq DNA polymerase, Mg2+, primer , dNTPs and template DNA. The results were as follows: DNA 60-150 ng, Taq DNA polymerase 1.0-1.5U, primer concentration 0.3-0.4 mmol/L, dNTPs concentration 200-250 μmol/L, Mg2+ concentration 2.5-3.0 mmol/L in 20μL PCR system. Under above optimal conditions,the abundant, stable and clear strips could be obtained.
刘丹,骆健美,刘峰,王敏. 褐黄孢链霉菌基因组DNA的提取及RAPD体系的优化[J]. 中国生物工程杂志, 2008, 28(专刊): 172-177.
. Genomic DNA extraction and optimization of RAPD amplifying conditions of Streptomyces gilvosporeus. China Biotechnology, 2008, 28(专刊): 172-177.
https://manu60.magtech.com.cn/biotech/CN/ 或 https://manu60.magtech.com.cn/biotech/CN/Y2008/V28/I专刊/172
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