活性污泥总DNA提取方法的研究

Extraction of DNA from Activated Sludge

采用冻融+玻璃珠、溶菌酶+SDS和冻融+玻璃珠+溶菌酶+SDS的方法提取活性污泥的微生物总DNA,并对以上三种方法进行比较,从中确定最佳的方法,以实现普通实验条件下成功提取符合PCR扩增要求的DNA。经紫外光度分析及电泳结果表明,冻融+玻璃珠+溶菌酶+SDS方法所得的DNA的OD260OD280为1.81,电泳结果显示,所提DNA片段分子量大于10kb,适于酶解和PCR扩增要求,为PCR技术应用于活性污泥的研究提供了一种简便、可靠的DNA提取方法。

In order to extract DNA from activated sludge under simple and easy experimental terms for follow- up PCR analysis, three methods（ freeze- thaw ＋ heads, lysozyme ＋ SDS, and freeze-thaw ＋ beads ＋ lysozyme ＋ SDS） were used and compared, results showed that the combination of freeze- thaw, lysozyme incubation and SDS lysis was the most efficient one. The OD260/OD280 of DNA sample was 1.81, and the result of electrophoresis showed that isolated DNA was over 10kb in size, which indicated that the extracted DNA could be used for PCR amplification. Thus, a rapid and less instrument - dependent protocol for direct extraction of DNA from activated sludge sample for PCR analysis was established.