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A method for obtaining DNA from compost

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Abstract

An effective cell lysis method for extraction of bacterial genomic DNA from compost was developed in this study. Enzymatic disruption method, physical–chemical combination method, and commercial kit method were used to extract DNA from compost samples and were compared by analyzing DNA yield and efficient cell lysis. The results showed that all the three methods can be used to extract high-quality DNA from compost, but the enzymatic method had better cell lysis efficiency and DNA yields than others without the use of special equipment and expensive spending. Comparison of different methods for lysing gram-positive bacteria Bacillus subtilis indicated that the enzymatic cell lysis is superior for destroying the gram-positive cell wall. Spin-bind DNA column was used for DNA purification, and the purity of the purified sample was checked by polymerase chain reaction to amplify a region of the 16S rRNA. Results indicated that the part of 16S rRNA were amplified from all the purified DNA samples, and all the amplification products could be digested by the restriction enzyme HhaI.

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Acknowledgments

This study was financially supported by the National Key Technology R&D Program of China (2006BAD14A05-02) and (2008BADB2B00); National 863 Projects (2007AA10Z166); National Natural Science Foundation (30800797)

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Correspondence to Changyan Deng.

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Wu, L., Li, F., Deng, C. et al. A method for obtaining DNA from compost. Appl Microbiol Biotechnol 84, 389–395 (2009). https://doi.org/10.1007/s00253-009-2103-8

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Keywords

  • Compost
  • Cell lysis
  • DNA extraction
  • 16S rRNA