水体中基因组提取方法的比较及水体中铜绿假单胞菌检测技术的研究

Comparison of the extraction methods of DNA and study on the biotechnological detection of Pseudomonas aeruginosa in river

以河水为研究对象,采用沸水浴法、SDS法、CTAB法、酚氯仿法和改良法提取河水中的基因组,对提取效果进行比较,确定出最佳提取方法;并利用传统培养法和分子生物学方法对人工污染样品中的铜绿假单胞菌进行检测,对两种方法检测结果进行比较;最后实验利用4种自然界水体验证了分子生物学方法的准确性。结果表明加入溶菌酶的酚氯仿法对河水中的DNA提取效果最好;传统培养法和分子生物学方法检测结果一致,且分子生物学方法比传统培养法节省一半左右的时间;用分子生物学方法可以检测出自然界水体中的铜绿假单胞菌。

Boiling-water method,SDS method,CTAB method,phenol-chloroform method and modified method were used to extract DNA from river,then the extracting results were compared to determine the best method.Traditional method and biotechnological method were used to detect Pseudomonas aeruginosa in samples after artificial contamination,then the results were compared.And the biotechnological method was used to detect 4 river samples in nature.The results showed that phenol-chloroform method with lysozyme was the best method to extract DNA from river.The same result was obtained when detecting the samples after artificial contamination using traditional method and biotechnological method,but the latter saved half the time spent in the former.The biotechnological method could be used to detect Pseudomonas aeruginosa from river in nature.