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DNA degradation in fish: Practical solutions and guidelines to improve DNA preservation for genomic research
The more demanding requirements of DNA preservation for genomic research can be difficult to meet when field conditions limit the methodological approaches that can be used or cause samples to be stored in suboptimal conditions. Such limitations may increase rates of DNA degradation, potentially ren...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7452763/ https://www.ncbi.nlm.nih.gov/pubmed/32884647 http://dx.doi.org/10.1002/ece3.6558 |
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author | Oosting, Tom Hilario, Elena Wellenreuther, Maren Ritchie, Peter A. |
author_facet | Oosting, Tom Hilario, Elena Wellenreuther, Maren Ritchie, Peter A. |
author_sort | Oosting, Tom |
collection | PubMed |
description | The more demanding requirements of DNA preservation for genomic research can be difficult to meet when field conditions limit the methodological approaches that can be used or cause samples to be stored in suboptimal conditions. Such limitations may increase rates of DNA degradation, potentially rendering samples unusable for applications such as genome‐wide sequencing. Nonetheless, little is known about the impact of suboptimal sampling conditions. We evaluated the performance of two widely used preservation solutions (1. DESS: 20% DMSO, 0.25 M EDTA, NaCl saturated solution, and 2. Ethanol >99.5%) under a range of storage conditions over a three‐month period (sampling at 1 day, 1 week, 2 weeks, 1 month, and 3 months) to provide practical guidelines for DNA preservation. DNA degradation was quantified as the reduction in average DNA fragment size over time (DNA fragmentation) because the size distribution of DNA segments plays a key role in generating genomic datasets. Tissues were collected from a marine teleost species, the Australasian snapper, Chrysophrys auratus. We found that the storage solution has a strong effect on DNA preservation. In DESS, DNA was only moderately degraded after three months of storage while DNA stored in ethanol showed high levels of DNA degradation already within 24 hr, making samples unsuitable for next‐generation sequencing. Here, we conclude that DESS was the most promising solution when storing samples for genomic applications. We recognize that the best preservation protocol is highly dependent on the organism, tissue type, and study design. We highly recommend performing similar experiments before beginning a study. This study highlights the importance of testing sample preservation protocols and provides both practical and economical advice to improve DNA preservation when sampling for genome‐wide applications. |
format | Online Article Text |
id | pubmed-7452763 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-74527632020-09-02 DNA degradation in fish: Practical solutions and guidelines to improve DNA preservation for genomic research Oosting, Tom Hilario, Elena Wellenreuther, Maren Ritchie, Peter A. Ecol Evol Original Research The more demanding requirements of DNA preservation for genomic research can be difficult to meet when field conditions limit the methodological approaches that can be used or cause samples to be stored in suboptimal conditions. Such limitations may increase rates of DNA degradation, potentially rendering samples unusable for applications such as genome‐wide sequencing. Nonetheless, little is known about the impact of suboptimal sampling conditions. We evaluated the performance of two widely used preservation solutions (1. DESS: 20% DMSO, 0.25 M EDTA, NaCl saturated solution, and 2. Ethanol >99.5%) under a range of storage conditions over a three‐month period (sampling at 1 day, 1 week, 2 weeks, 1 month, and 3 months) to provide practical guidelines for DNA preservation. DNA degradation was quantified as the reduction in average DNA fragment size over time (DNA fragmentation) because the size distribution of DNA segments plays a key role in generating genomic datasets. Tissues were collected from a marine teleost species, the Australasian snapper, Chrysophrys auratus. We found that the storage solution has a strong effect on DNA preservation. In DESS, DNA was only moderately degraded after three months of storage while DNA stored in ethanol showed high levels of DNA degradation already within 24 hr, making samples unsuitable for next‐generation sequencing. Here, we conclude that DESS was the most promising solution when storing samples for genomic applications. We recognize that the best preservation protocol is highly dependent on the organism, tissue type, and study design. We highly recommend performing similar experiments before beginning a study. This study highlights the importance of testing sample preservation protocols and provides both practical and economical advice to improve DNA preservation when sampling for genome‐wide applications. John Wiley and Sons Inc. 2020-07-13 /pmc/articles/PMC7452763/ /pubmed/32884647 http://dx.doi.org/10.1002/ece3.6558 Text en © 2020 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Original Research Oosting, Tom Hilario, Elena Wellenreuther, Maren Ritchie, Peter A. DNA degradation in fish: Practical solutions and guidelines to improve DNA preservation for genomic research |
title | DNA degradation in fish: Practical solutions and guidelines to improve DNA preservation for genomic research |
title_full | DNA degradation in fish: Practical solutions and guidelines to improve DNA preservation for genomic research |
title_fullStr | DNA degradation in fish: Practical solutions and guidelines to improve DNA preservation for genomic research |
title_full_unstemmed | DNA degradation in fish: Practical solutions and guidelines to improve DNA preservation for genomic research |
title_short | DNA degradation in fish: Practical solutions and guidelines to improve DNA preservation for genomic research |
title_sort | dna degradation in fish: practical solutions and guidelines to improve dna preservation for genomic research |
topic | Original Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7452763/ https://www.ncbi.nlm.nih.gov/pubmed/32884647 http://dx.doi.org/10.1002/ece3.6558 |
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