一种简单、有效的适于PCR操作的放线菌DNA提取方法

A Quick and Efficient Method for Genomic DNA Extraction from Actionbacteria

目的:利用改良酶法发展了一种从微量(几百微升)发酵液中快速安全的提取放线菌基因组DNA的方法。方法:利用溶菌酶破壁,蛋白酶K和SDS除蛋白,成功提取较高质量的放线菌基因组DNA,所得的DNA可作为PCR反应的模板进行16SrRNA等基因有效扩增。结果:能从海绵和土壤分离的放线菌中成功提取基因组DNA。结论:该方法操作简单、费用低廉、不使用酚、氯仿等有毒害作用有机试剂,非常适于长期从事放线菌操作的研究人员。为大量放线菌菌株的快速鉴别、高通量筛选和系统分类研究创造了条件。

Objective：A brief and safe method of extracting genomic DNA from small-scale（hundred microlitres） actinobacterial fermentation was developed.Method： Lysozyme was used to lysis cell wall and proteinase K and SDS was used to remove protein.Conclusion： Genomic DNA was extracted successfully from actinobacteria isolated from marine sponge and soil;The DNA is suitable for PCR amplification.The method can be used in actinobacteria systematics and rapid identification.