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Methods for extraction of microorganism DNA from glacier surface snow

ISSN号：1001-7410
期刊名称：《第四纪研究》
时间：0
分类：S792.14[农业科学—林木遗传育种;农业科学—林学] X703[环境科学与工程—环境工程]
作者机构：[1]State Key Laboratory of Cryospheric Sciences, Cold and Arid Regions Environmental and Engineering Research Institute, Chinese Academy of Sciences, Lanzhou, Gansu 730000, China, [2]School of Geographic and Oceanographic Sciences, Nanjing University, Nanjing, Jiangsu 210093, China, [3]Shangqiu Normal University, Department of Life Science, Shangqiu, Henan 476000, China
相关基金：This research was supported by the National Natural Science Fotmdation of China （Nos. 40825017, 40576001 and 31100369）.

作者： PeiYing Yan[1], ShuGui Hou[1,2], Tuo Chen[1], ShuHong Zhang[3], WeiJun Sun[1]

关键词： DNA提取方法, 微生物, 冰川, 表面, 积雪, DNA提取法, 过滤膜, 蛋白酶K, glacier surface snow, extraction of microorganism DNA, optimizing method

中文摘要：

In order to thoroughly investigate the diversity of glacier microorganisms, four DNA extraction methods with different lysis patterns were tested and two screened methods (the Bosshard-Bano method and the Zhou method) were optimized for the most effective 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 from glacier surface snow. Procedurally, it was found that a modified Bosshard-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.

英文摘要：

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.