2014
DOI: 10.1111/1755-0998.12281
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The room temperature preservation of filtered environmental DNA samples and assimilation into a phenol–chloroform–isoamyl alcohol DNA extraction

Abstract: Current research targeting filtered macrobial environmental DNA (eDNA) often relies upon cold ambient temperatures at various stages, including the transport of water samples from the field to the laboratory and the storage of water and/or filtered samples in the laboratory. This poses practical limitations for field collections in locations where refrigeration and frozen storage is difficult or where samples must be transported long distances for further processing and screening. This study demonstrates the s… Show more

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Cited by 301 publications
(385 citation statements)
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“…DNA was isolated following a modified chloroform -isoamyl alcohol (24:1; Amresco) extraction and an isopropanol precipitation. (Renshaw et al 2015; see full details in the Supplementary material, Appendix S1 1 ). To remove potential inhibitors, resuspended DNA was treated with the OneStepPCR Inhibitor Removal Kit (Zymo Research, Irvine, California, USA).…”
Section: Sample Processing and Extractionmentioning
confidence: 99%
“…DNA was isolated following a modified chloroform -isoamyl alcohol (24:1; Amresco) extraction and an isopropanol precipitation. (Renshaw et al 2015; see full details in the Supplementary material, Appendix S1 1 ). To remove potential inhibitors, resuspended DNA was treated with the OneStepPCR Inhibitor Removal Kit (Zymo Research, Irvine, California, USA).…”
Section: Sample Processing and Extractionmentioning
confidence: 99%
“…We kept these samples on ice until they could be processed in the lab (within hours of collection). We filtered the total volume of the samples (1 L) onto cellulose acetate filters (47 mm diameter; 0.45 um pore size) under vacuum pressure, and preserved the filter at room temperature in Longmire's buffer following (Renshaw et al, 2015). Deionized water (1-L) served as a negative control for filtering.…”
Section: Environmental Setting and Field Collectionsmentioning
confidence: 99%
“…Deionized water (1-L) served as a negative control for filtering. We extracted total DNA from the filters using the phenol:chloroform:isoamyl alcohol protocol in Renshaw et al (2015), resuspended the eluate in 200 uL water, and used 1 uL of diluted DNA extract (1:100) as template for PCR. See Supplemental Methods for additional sampling details.…”
Section: Environmental Setting and Field Collectionsmentioning
confidence: 99%
“…We used a standard cetyl trimethyl ammonium bromide (CTAB) extraction method that has been used in numerous eDNA studies [1,28,36] and verified by Renshaw et al [38]. Therefore, capture efficiency should not have been a concern.…”
Section: Dna Extractionmentioning
confidence: 99%