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An Extremely Simple and Effective Colony PCR Procedure for Bacteria, Yeasts, and Microalgae

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Abstract

An extremely simple and effective colony PCR procedure is established for both gram-negative and gram-positive bacteria, yeasts, and microalgae. Among the four lysis buffers examined, Y-PER is observed to be more effective than Tris/EDTA, 0.2 % SDS, and 10 mM EDTA in the extraction of PCR-quality genomic DNA from those microorganisms. Vortexing or pipetting agitation of the cells in Y-PER for 5–10 s was sufficient to release genomic DNA for all the test bacteria and yeasts, and most microalgae. Additional incubation at 98 °C for 5 min for further cell disruption was essential only for Chlorella vulgaris due to its notoriously rigid cell wall.

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Acknowledgments

We thank the IB group for their insights and discussion. This work was funded by A*STAR in Singapore ICES/12-574A01.

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Authors and Affiliations

  1. Mathematisch-Naturwissenschaftliche Fakultät, Rheinische Friedrich-Wilhelms Universität Bonn, Wegelerstraße 10, 53115, Bonn, Germany

    Heiko Packeiser

  2. Industrial Biotechnology Program, Institute of Chemical & Engineering Sciences, Agency for Science, Technology and Research (A*STAR), 1 Pesek Road, Jurong Island, 627833, Singapore, Singapore

    Chanyuen Lim, Balaji Balagurunathan, Jinchuan Wu & Hua Zhao

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  1. Heiko Packeiser
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  2. Chanyuen Lim
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  3. Balaji Balagurunathan
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  4. Jinchuan Wu
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  5. Hua Zhao
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Corresponding author

Correspondence to Hua Zhao.

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Packeiser, H., Lim, C., Balagurunathan, B. et al. An Extremely Simple and Effective Colony PCR Procedure for Bacteria, Yeasts, and Microalgae. Appl Biochem Biotechnol 169, 695–700 (2013). https://doi.org/10.1007/s12010-012-0043-8

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  • DOI: https://doi.org/10.1007/s12010-012-0043-8

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