Optimization of Extracting Method for Microbial Genome DNA in Coal Geological Environment

doi:10.13560/j.cnki.biotech.bull.1985.2017-1145

Abstract

Abstract: High-quality DNA is the basis for molecular biology research. By improving the traditional method of extracting DNA（phenol-chloroform method）and optimizing the cell disruption conditions while having Rhodococcus sp.R04 and pulverized coal as experimental materials,a modified method of efficiently extracting genomic DNA from microbes in coal geological environment has been established. The microbial genomic DNAs extracted by this modified method and the commercial kit were evaluated by agarose gel electrophoresis and PCR amplifications of bacterial and archaea specific fragments. The results showed that the microbial genome DNAs from coal geological environment were obtained by both the modified method and the kit method,and used as temple for various specific PCR amplifications. Compared with the extracted DNA with the commercial kit,the DNA by the modified method had obvious main band,accounted about 50% of total DNA content,and the molecular weight of the DNA was nearly 23 kb. Also,the extraction amount of DNA was very large,and was about 5-10 times that by the commercial kit. Moreover,DNA by the modified method can be used in many purposes such as DNA library construction and metagenomic sequencing. Consequently,the modified method for extracting microbial genomic DNA from coal geological environment is not only cheap and effective,also the extracted DNA is in high-quality,thus it is very suitable for laboratory and scientific researches.

Key words: microorganism in coal geological environment, genomic DNA extraction, DNA quality, PCR amplification

YANG Xiu-qing, CHEN Yan-mei, WU Rui-wei, WANG Bao-yu, HAN Zuo-ying. Optimization of Extracting Method for Microbial Genome DNA in Coal Geological Environment[J]. Biotechnology Bulletin, 2018, 34(9): 177-183.

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