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KMID : 0880220150530110767
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Journal of Microbiology
2015 Volume.53 No. 11 p.767 ~ p.775
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Enhanced method for microbial community DNA extraction and purification from agricultural yellow loess soil
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Kathiravan Mathur Nadarajan
Gim Geun-Ho
Ryu Jae-Won
Kim Pyung-Il
Lee Chul-Won
Kim Si-Wouk
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Abstract
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In this study, novel DNA extraction and purification methods were developed to obtain high-quantity and reliable quality DNA from the microbial community of agricultural yellow loess soil samples. The efficiencies of five different soil DNAextraction protocols were evaluated on the basis of DNA yield, quality and DNA shearing. Our suggested extraction method, which used CTAB, EDTA and cell membrane lytic enzymes in the extraction followed by DNA precipitation using isopropanol, yielded a maximum DNA content of 42.28 ± 5.59 μg/g soil. In addition, among the five different purification protocols, the acid-treated polyvinyl polypyrrolidone (PVPP) spin column purification method yielded high-quality DNA and recovered 91% of DNA from the crude DNA. Spectrophotometry revealed that the ultraviolet A 260/A 230 and A 260/A 280 absorbance ratios of the purified DNA were 1.82 ± 0.03 and 1.94 ± 0.05, respectively. PCR-based 16S rRNA amplification showed clear bands at ~1.5 kb with acid-treated PVPP?purified DNA templates. In conclusion, our suggested extraction and purification protocols can be used to recover high concentration, high purity, and high-molecular-weight DNA from clay and silica-rich agricultural soil samples.
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KEYWORD
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microbial community DNA, metagenomics, soil texture, DNA extraction purification
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