The room temperature preservation of filtered environmental DNA samples and assimilation into a phenol-chloroform-isoamyl alcohol DNA extraction.
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 successful preservation of eDNA at room temperature (20 °C) in two lysis buffers, CTAB and Longmire's, over a 2-week period of time. Moreover, the preserved eDNA samples were seamlessly integrated into a phenol-chloroform-isoamyl alcohol (PCI) DNA extraction protocol. The successful application of the eDNA extraction to multiple filter membrane types suggests the methods evaluated here may be broadly applied in future eDNA research. Our results also suggest that for many kinds of studies recently reported on macrobial eDNA, detection probabilities could have been increased, and at a lower cost, by utilizing the Longmire's preservation buffer with a PCI DNA extraction.
Department of Biological Sciences, University of Notre Dame, 100 Galvin Life Sciences Center, Notre Dame, IN, 46556, USA.
Mark A
MA
Renshaw
Brett P
BP
Olds
Christopher L
CL
Jerde
Margaret M
MM
McVeigh
David M
DM
Lodge
D000438:Alcohols; D004247:DNA / Q000302:isolation & purification*; D005374:Filtration / Q000379:methods*; D056186:Metagenomics / Q000379:methods*; D019800:Phenol; D011309:Preservation, Biological / Q000379:methods*; D013696:Temperature
D000438:Alcohols; D019800:Phenol; D004247:DNA
Lepomis macrochirus; eDNA extraction; eDNA preservation; environmental DNA