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Recommendations for the processing and storage of water samples before polymerase chain reaction (PCR) analysis
Authors:BJ Gilpin  M Devane  F Nourozi  B Robson  P Scholes  S Lin
Institution: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
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.
Keywords:microbial source tracking  water quality  DNA storage  qPCR  human-specific Bacteroidales
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