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An improved protocol for the isolation of total genomic DNA from Labyrinthulomycetes

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Abstract

Many protocols have been used for extraction of DNA from Thraustochytrids. These generally involve the use of CTAB, phenol/chloroform and ethanol. They also feature mechanical grinding, sonication, N2 freezing or bead beating. However, the resulting chemical and physical damage to extracted DNA reduces its quality. The methods are also unsuitable for large numbers of samples. Commercially-available DNA extraction kits give better quality and yields but are expensive. Therefore, an optimized DNA extraction protocol was developed which is suitable for Thraustochytrids to both minimise expensive and time-consuming steps prior to DNA extraction and also to improve the yield. The most effective method is a combination of single bead in TissueLyser (Qiagen) and Proteinase K. Results were conclusive: both the quality and the yield of extracted DNA were higher than with any other method giving an average yield of 8.5 µg/100 mg biomass.

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Acknowledgments

We would like to thank Dr. Tom Lewis (Tasmania), Solaris Bioscience (Tasmania) and Queensland University of Technology (Queensland) for their support of this project.

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Authors and Affiliations

  1. Faculty of Health, Queensland University of Technology, 02 George Street, Brisbane, QLD, 4001, Australia

    Chaminda Padmashantha Ranasinghe & Megan Hargreaves

  2. Centre for Tropical Crops and Biocommodities, Science and Engineering Faculty, Queensland University of Technology, 02 George Street, Brisbane, QLD, 4001, Australia

    Rob Harding

Authors
  1. Chaminda Padmashantha Ranasinghe
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  2. Rob Harding
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  3. Megan Hargreaves
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Correspondence to Chaminda Padmashantha Ranasinghe.

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Ranasinghe, C.P., Harding, R. & Hargreaves, M. An improved protocol for the isolation of total genomic DNA from Labyrinthulomycetes. Biotechnol Lett 37, 685–690 (2015). https://doi.org/10.1007/s10529-014-1712-1

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  • DOI: https://doi.org/10.1007/s10529-014-1712-1

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