Rapid, high-throughput extraction of bacterial genomic DNA from selective-enrichment culture media |
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Authors: | Wilson T Carson J |
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Affiliation: | Fish Health Unit, Tasmanian Aquaculture and Fisheries Institute, University of Tasmania, Launceston, Australia. Teresa.Wilson@dpiwe.tas.gov.au |
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Abstract: | AIMS: To create a fast, sensitive and inexpensive high-throughput method for the extraction of bacterial genomic DNA from selective-enrichment culture media. METHODS AND RESULTS: Lysis of bacteria was achieved using guanidinium isothiocyanate, and DNA was extracted using 96-well glass microfibre filtration plates. Extraction-PCR detected the presence of 1 cfu Yersinia ruckeri and 16 cfu Lactococcus garvieae 200 microl(-1) sample of selective-enrichment medium. CONCLUSION: An efficient method for high-throughput extraction of bacterial genomic DNA from selective-enrichment culture media was achieved. SIGNIFICANCE AND IMPACT OF THE STUDY: This method enables detection of covert bacterial infections in fish. The simultaneous extraction of large numbers of samples allows for its use in bacterial monitoring programmes and quarantine. |
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