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No filters, no fridges: a method for preservation of water samples for eDNA analysis

BACKGROUND: Advancements in the detection of environmental DNA (eDNA) for detecting species of interest will likely allow for expanded use of these techniques in the field. One obstacle that continues to hinder applications in the field is the requirement of a cold chain of storage for water samples...

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Autores principales: Williams, Kelly E., Huyvaert, Kathryn P., Piaggio, Antoinette J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4898389/
https://www.ncbi.nlm.nih.gov/pubmed/27278936
http://dx.doi.org/10.1186/s13104-016-2104-5
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author Williams, Kelly E.
Huyvaert, Kathryn P.
Piaggio, Antoinette J.
author_facet Williams, Kelly E.
Huyvaert, Kathryn P.
Piaggio, Antoinette J.
author_sort Williams, Kelly E.
collection PubMed
description BACKGROUND: Advancements in the detection of environmental DNA (eDNA) for detecting species of interest will likely allow for expanded use of these techniques in the field. One obstacle that continues to hinder applications in the field is the requirement of a cold chain of storage for water samples containing eDNA. While eDNA has been successfully preserved using Longmire’s lysis buffer applied to filters, it has yet to be tried with freshwater samples collected for eDNA detection of an invasive species. We tested the utility of Longmire’s solution (100 mM Tris, 100 mM EDTA, 10 mM NaCl, 0.5 % SDS, 0.2 % sodium azide) as an additive to freshwater samples for preservation of eDNA. RESULTS: Environmental DNA was effectively preserved in 15 mL water samples with Longmire’s solution added; eDNA positive detection was comparable to freezing the samples at −80 °C and occurred out to 56 days at the highest concentration (5 mL Longmire’s solution: 15 mL sample water). Medium and low concentrations of Longmire’s solution added to 15 mL of sample water generally preserved eDNA out to 56 days but not as well as did freezing or application of the highest concentration of Longmire’s lysis buffer. Treatment and degradation time had a significant effect on average DNA concentration of samples, although not the interaction of treatment and time. Perfect detection occurred out to 56 days with the high Longmire’s treatment group but DNA concentration was significantly lower at this time point compared to 28 days. CONCLUSION: We conclude that Longmire’s lysis buffer is a viable alternative to cold chain storage that can simplify the collection of eDNA by eliminating the need for filtering and allow more time for sample collection when added at our highest concentration (1 part Longmire’s:3 parts water sample), which could translate to an increase in the chances of detecting a rare or elusive species. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13104-016-2104-5) contains supplementary material, which is available to authorized users.
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spelling pubmed-48983892016-06-09 No filters, no fridges: a method for preservation of water samples for eDNA analysis Williams, Kelly E. Huyvaert, Kathryn P. Piaggio, Antoinette J. BMC Res Notes Technical Note BACKGROUND: Advancements in the detection of environmental DNA (eDNA) for detecting species of interest will likely allow for expanded use of these techniques in the field. One obstacle that continues to hinder applications in the field is the requirement of a cold chain of storage for water samples containing eDNA. While eDNA has been successfully preserved using Longmire’s lysis buffer applied to filters, it has yet to be tried with freshwater samples collected for eDNA detection of an invasive species. We tested the utility of Longmire’s solution (100 mM Tris, 100 mM EDTA, 10 mM NaCl, 0.5 % SDS, 0.2 % sodium azide) as an additive to freshwater samples for preservation of eDNA. RESULTS: Environmental DNA was effectively preserved in 15 mL water samples with Longmire’s solution added; eDNA positive detection was comparable to freezing the samples at −80 °C and occurred out to 56 days at the highest concentration (5 mL Longmire’s solution: 15 mL sample water). Medium and low concentrations of Longmire’s solution added to 15 mL of sample water generally preserved eDNA out to 56 days but not as well as did freezing or application of the highest concentration of Longmire’s lysis buffer. Treatment and degradation time had a significant effect on average DNA concentration of samples, although not the interaction of treatment and time. Perfect detection occurred out to 56 days with the high Longmire’s treatment group but DNA concentration was significantly lower at this time point compared to 28 days. CONCLUSION: We conclude that Longmire’s lysis buffer is a viable alternative to cold chain storage that can simplify the collection of eDNA by eliminating the need for filtering and allow more time for sample collection when added at our highest concentration (1 part Longmire’s:3 parts water sample), which could translate to an increase in the chances of detecting a rare or elusive species. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13104-016-2104-5) contains supplementary material, which is available to authorized users. BioMed Central 2016-06-08 /pmc/articles/PMC4898389/ /pubmed/27278936 http://dx.doi.org/10.1186/s13104-016-2104-5 Text en © The Author(s) 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Technical Note
Williams, Kelly E.
Huyvaert, Kathryn P.
Piaggio, Antoinette J.
No filters, no fridges: a method for preservation of water samples for eDNA analysis
title No filters, no fridges: a method for preservation of water samples for eDNA analysis
title_full No filters, no fridges: a method for preservation of water samples for eDNA analysis
title_fullStr No filters, no fridges: a method for preservation of water samples for eDNA analysis
title_full_unstemmed No filters, no fridges: a method for preservation of water samples for eDNA analysis
title_short No filters, no fridges: a method for preservation of water samples for eDNA analysis
title_sort no filters, no fridges: a method for preservation of water samples for edna analysis
topic Technical Note
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4898389/
https://www.ncbi.nlm.nih.gov/pubmed/27278936
http://dx.doi.org/10.1186/s13104-016-2104-5
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