Comparison of Methods for Total DNA Extraction of Microorganisms in Glacier Surface Snow Made Progress
Updatetime:2013-09-25From:
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Microorganisms deposit in glacier with the atmospheric circulation and precipitation, and they are main life forms in extreme low temperature environment. Microorganisms in glacier make an effect to the surface albedo of glacier and the hydrochemistry; they play an important role in producing the glacier in the primary stage and the circulation of materials and energy flow. With traditional cultured methods, the research on microorganisms in glacier focuses on the structure and the diversity of microflora in the ice core and the nivation cirque in different regions. However, cultured microorganisms may be less than 1% of the total number of all microorganisms in natural environment, which limit the environmental microbiology research.
We used the molecular biology methods of the molecular hybridization, DGGE (TGGE) and the high-throughput sequencing to study the number of environmental microbiology, the characters of community structure and its functions through the culture-independent in the mid-1980s. The premise of these technologies is DNA extraction of microorganisms.
The science and research personnel in CAREERI screened out a better method through comparison of the four methods for total DNA extraction of microorganisms with different lysis. The method screened out was used to optimize the extraction and precipitation. Finally, the optimized method was compared with commercial kit for total DNA extraction of microorganisms. The results showed that the commercial kit was fit for total DNA extraction of microorganisms. The results showed that the commercial kit was fit for total DNA extraction of microorganisms in surface snow on glacier, and another method with cutting the filter membrane into pieces, lysis (lysozyme (5mg•mL-1)+protease K (1mg•mL-1)+CTAB(1%)+SDS(1%)), extraction once by chloroform-isoamylalcohol (24:1) and DNA precipitation by ethanol, was an effective and low – cost method for total DNA extraction of microorganisms in surface snow on glacier.
This research is of important significance to the molecular polymorphism of microorganisms in glacier and its effects on geo-biochemical circulation.
This program was funded by the National Natural Science Foundation of China (40825017, 40576001, 31100369). And the research achievement published in the Journal of Glaciology and Geocryology
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