Recommendations for the processing and storage of water samples before polymerase chain reaction (PCR) analysis |
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Authors: | BJ Gilpin M Devane F Nourozi B Robson P Scholes S Lin |
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Affiliation: | 1. Christchurch Science Centre, Institute of Environmental Science and Research (ESR) Ltd, Christchurch New Zealandbrent.gilpin@esr.cri.nz;3. Christchurch Science Centre, Institute of Environmental Science and Research (ESR) Ltd, Christchurch New Zealand |
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Abstract: | A range of options for the storage and partial processing of water samples before polymerase chain reaction (PCR) analysis were evaluated. In addition, temporal storage of extracted DNA at 4 °C was investigated. Filtering the water sample and then storing that filter at ?20 °C for up to six months before DNA extraction was equivalent to extracting DNA immediately. Freezing a water sample resulted in an immediate 1 log loss of PCR signal, although thereafter the sample was stable for at least three months. A rapid loss of detectable PCR product was found when storing a water sample at 4 °C for longer than one week. Extracted DNA could be stored at 4 °C for at least six months without considerable loss of PCR signal. |
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Keywords: | microbial source tracking water quality DNA storage qPCR human-specific Bacteroidales |
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