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A simplified quick microbial genomic DNA extraction via freeze–thawing cycles

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Abstract

Effective isolation of high-quality genomic DNA is one of the essential steps in molecular biology, biochemistry, and genetic studies. Here we describe a simplified procedure based on repeated freeze–thawing cycles to isolate genomic DNA from different organisms of microbes (Burkholderia pyrrocinia JK-SH007, Bacillus pumilus HRl0, Botrytis cinerea) and nematodes (Bursaphelenchus xylophilus). The DNA extraction buffer includes 10% of CTAB; 4% of NaCl (W/V); 20 mM of ethylenediamine tetraacetic acid; 100 mM of Tris-HCl, pH 8.0 and 1% of polyvinylpyrrolidone. The released DNA was purified from the mixture using a phenol/chloroform mixture and precipitated in 70% ethanol to remove proteins, carbohydrates, phenols, RNA, etc. Our method is a reproducible, simple, and rapid technique for routine DNA extractions from various microorganisms and nematodes. Furthermore, the low cost of this method could be an economic benefit to large-scale studies.

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Acknowledgements

This work was financially supported by the Jiangsu Agricultural Science and Technology Independent Innovation Fund (CX16-1005) and the Chinese State Forestry Administration Special Research Program for Forestry Sectors Beneficial to the Public (No. 201304404). This work was also financially supported by the Shanghai Science and Technology Agriculture Key Project (2014:5-6) and the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD).

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Authors and Affiliations

  1. College of Forestry and Co-Innovation Center for Sustainable Forestry in Southern China, Nanjing Forestry University, 159 Longpan Road, Nanjing, Jiangsu, 210037, China

    Feifei Chen, Jianren Ye, Wanhui Liu & Jiyuan Shi

  2. Key Laboratory for Prevention and Management of Invasive Species, Nanjing Forestry University, Nanjing, Jiangsu, 210037, China

    Feifei Chen, Jianren Ye, Wanhui Liu & Jiyuan Shi

  3. Department of Biology, Lakehead University, Thunder Bay, ON, P7B 5E1, Canada

    Feifei Chen, Chonlong Chio & Wensheng Qin

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  1. Feifei Chen
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  2. Jianren Ye
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  3. Chonlong Chio
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  4. Wanhui Liu
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Correspondence to Jianren Ye.

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Chen, F., Ye, J., Chio, C. et al. A simplified quick microbial genomic DNA extraction via freeze–thawing cycles. Mol Biol Rep 47, 703–709 (2020). https://doi.org/10.1007/s11033-019-05176-w

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