Abstract
Current DNA extraction methods for parasites are labour-intensive and usually involve several steps, increasing the potential for cross-contamination. We describe here a closed-tube DNA extraction procedure based upon the use of a thermostable proteinase that enabled sensitive amplification of target loci from parasites from diverse lineages including Apicomplexa, Sarcomastgophora and Nematoda. Moreover, this procedure is not subject to cross-contamination and is readily adaptable to automation.
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Acknowledgements
This research was funded by the Environmental Biotechnology CRC Pty Ltd., a centre established and supported under the Australian Government’s Cooperative Research Centres Program. The authors would like to thank ZyGEM Corp. Ltd. for the supply of prepGEM for evaluation, Dr Rogan Lee from Westmead Hospital, Sydney, for the supply of human faecal samples, Dr Simon Reid (Murdoch University) for provision of Giardia trophozoites and Prof Nick Sangster (Sydney University) for supply of nematode eggs, adults and larvae.
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Ferrari, B.C., Power, M.L. & Bergquist, P.L. Closed-tube DNA extraction using a thermostable proteinase is highly sensitive, capable of single parasite detection. Biotechnol Lett 29, 1831–1837 (2007). https://doi.org/10.1007/s10529-007-9487-2
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DOI: https://doi.org/10.1007/s10529-007-9487-2