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      DNA degradation in fish: Practical solutions and guidelines to improve DNA preservation for genomic research

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          Abstract

          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.

          Abstract

          The initial stages of DNA preservation can be crucial for obtaining high‐quality data for genomic research. However, environmental conditions in the field and during transport are typically suboptimal for preserving DNA, potentially spoiling costly samples. Here, we evaluate two cost‐effective preservation solutions and provide a comprehensive background on what causes DNA to degrade and practical guidelines on how to ensure the best approach for preserving DNA in field‐collected samples.

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          Most cited references25

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          Universal and rapid salt-extraction of high quality genomic DNA for PCR-based techniques.

          A very simple, fast, universally applicable and reproducible method to extract high quality megabase genomic DNA from different organisms is described. We applied the same method to extract high quality complex genomic DNA from different tissues (wheat, barley, potato, beans, pear and almond leaves as well as fungi, insects and shrimps' fresh tissue) without any modification. The method does not require expensive and environmentally hazardous reagents and equipment. It can be performed even in low technology laboratories. The amount of tissue required by this method is approximately 50-100 mg. The quantity and the quality of the DNA extracted by this method is high enough to perform hundreds of PCR-based reactions and also to be used in other DNA manipulation techniques such as restriction digestion, Southern blot and cloning.
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            Preservation of avian blood and tissue samples for DNA analyses

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              The Third Revolution in Sequencing Technology.

              Forty years ago the advent of Sanger sequencing was revolutionary as it allowed complete genome sequences to be deciphered for the first time. A second revolution came when next-generation sequencing (NGS) technologies appeared, which made genome sequencing much cheaper and faster. However, NGS methods have several drawbacks and pitfalls, most notably their short reads. Recently, third-generation/long-read methods appeared, which can produce genome assemblies of unprecedented quality. Moreover, these technologies can directly detect epigenetic modifications on native DNA and allow whole-transcript sequencing without the need for assembly. This marks the third revolution in sequencing technology. Here we review and compare the various long-read methods. We discuss their applications and their respective strengths and weaknesses and provide future perspectives.
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                Author and article information

                Contributors
                Tom.oosting@gmail.com
                Journal
                Ecol Evol
                Ecol Evol
                10.1002/(ISSN)2045-7758
                ECE3
                Ecology and Evolution
                John Wiley and Sons Inc. (Hoboken )
                2045-7758
                13 July 2020
                August 2020
                : 10
                : 16 ( doiID: 10.1002/ece3.v10.16 )
                : 8643-8651
                Affiliations
                [ 1 ] School of Biological Sciences Victoria University of Wellington Wellington New Zealand
                [ 2 ] The New Zealand Institute for Plant & Food Research Ltd Auckland New Zealand
                [ 3 ] Nelson Seafood Research Unit The New Zealand Institute for Plant & Food Research Ltd Nelson New Zealand
                [ 4 ] School of Biological Sciences The University of Auckland Auckland New Zealand
                Author notes
                [*] [* ] Correspondence

                Tom Oosting, School of Biology, Victoria University of Wellington, Kelburn Parade, 6012 Wellington, New Zealand.

                Email: Tom.oosting@ 123456gmail.com

                Author information
                https://orcid.org/0000-0002-7031-0747
                https://orcid.org/0000-0001-8124-8881
                https://orcid.org/0000-0002-2764-8291
                https://orcid.org/0000-0002-8351-7931
                Article
                ECE36558
                10.1002/ece3.6558
                7452763
                32884647
                886ffa7a-c52b-4aed-a856-a432fd185d29
                © 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.

                History
                : 24 March 2020
                : 26 May 2020
                : 10 June 2020
                Page count
                Figures: 3, Tables: 2, Pages: 9, Words: 6307
                Funding
                Funded by: Royal Marsden NHS Foundation Trust , open-funder-registry 10.13039/100012139;
                Award ID: 16‐VUW‐040
                Categories
                Original Research
                Original Research
                Custom metadata
                2.0
                August 2020
                Converter:WILEY_ML3GV2_TO_JATSPMC version:5.8.8 mode:remove_FC converted:27.08.2020

                Evolutionary Biology
                dmso,dna preservation,ethanol,fish,next‐generation sequencing,snapper
                Evolutionary Biology
                dmso, dna preservation, ethanol, fish, next‐generation sequencing, snapper

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