In order to thoroughly investigate the diversity of glacier microorganisms, four DNA extraction methods with differem lysis pat- terns were tested and two screened methods （the Bosshard-Bano method and the Zhou metho...In order to thoroughly investigate the diversity of glacier microorganisms, four DNA extraction methods with differem lysis pat- terns were tested and two screened methods （the Bosshard-Bano method and the Zhou method） were optimized for the most ef- fective form of the filter membrane （cut vs. uncut）, the DNA extraction method, and the precipitation method. The two optimized methods were then compared with the commercial Mo-Bio DNA extraction kit, and the results showed that the kit was generally suitable for extraction of microorganism DNA fi＇om glacier surface snow. Procedurally, it was found that a modified Boss- hard-Bano method （i.e., cutting the filter membrane into pieces, using a specific lysis pattern [lysozyme （5 mg/mL）-protease K （ 1 mg/mL）-CTAB （ 1%）-SDS （ 1%）], performing the extraction only once by chloroform-isoamyl alcohol （24： 1）, and conducting DNA precipitation by pure ethanol） was also an effective and less expensive method for extraction of microorganism DNA from glacier surface snow.展开更多
基金supported by the National Natural Science Foundation of China (Nos. 40825017, 40576001 and 31100369)
文摘In order to thoroughly investigate the diversity of glacier microorganisms, four DNA extraction methods with differem lysis pat- terns were tested and two screened methods （the Bosshard-Bano method and the Zhou method） were optimized for the most ef- fective form of the filter membrane （cut vs. uncut）, the DNA extraction method, and the precipitation method. The two optimized methods were then compared with the commercial Mo-Bio DNA extraction kit, and the results showed that the kit was generally suitable for extraction of microorganism DNA fi＇om glacier surface snow. Procedurally, it was found that a modified Boss- hard-Bano method （i.e., cutting the filter membrane into pieces, using a specific lysis pattern [lysozyme （5 mg/mL）-protease K （ 1 mg/mL）-CTAB （ 1%）-SDS （ 1%）], performing the extraction only once by chloroform-isoamyl alcohol （24： 1）, and conducting DNA precipitation by pure ethanol） was also an effective and less expensive method for extraction of microorganism DNA from glacier surface snow.
