An Improved Method for Extraction of Soil Microbial DNA for Metagenomics Study
Keywords:
16S rDNA, Humic Acidmetagenomics, Soil Microbial DNAAbstract
Metagenomics enables the genome wide study of unculturable microorganisms in any habitat. Standardization of microbial DNA isolation protocol is the first and foremost step for molecular characterization of unculturable microbes, DNA isolation methods from soil needs to be standardized. During the last 10 years, a number of protocols for DNA extraction from environmental samples have been reported. A number of methods are being used for lysis of cells in soil samples. In the present study, four different methods (i) by using both CTAB and SDS,3 (ii) only CTAB,4 (iii) enzymatic (lysozyme) lysis,5 and (iv) sonication6 for different time duration were used. It was found that the first three methods yielded very less amount of DNA and the quality ratio (A260/280) was also low, which indicated impurity in DNA. Genomic DNA isolation protocol of community DNA was optimized with sonication (30 sec) and modifications made in standard CTAB method4 and good quality and considerably higher yield was obtained in the method involving sonication followed by extraction with CTAB and SDS in the present study.
How to cite this article: Sharma N, Singh G, Sudarsan Y. An Improved Method for Extraction of Soil Microbial DNA for Metagenomics Study. J Adv Res Agri Sci Tech 2019; 2(2): 1-4.
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