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Development of a Rapid and Efficient Method for Non-Lethal DNA Sampling and Genotyping in Scallops

Non-lethal DNA sampling has long appealed to researchers studying population and conservation genetics, as it does not necessitate removing individuals permanently from their natural environment or destroying valuable samples. However, such an approach has not yet been well established in bivalves....

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Autores principales: Mao, Junxia, Lv, Jia, Miao, Yan, Sun, Changsen, Hu, Liping, Zhang, Ru, Fu, Xiaoteng, Zhang, Lingling, Hu, Xiaoli, Wang, Shi, Bao, Zhenmin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3706602/
https://www.ncbi.nlm.nih.gov/pubmed/23874509
http://dx.doi.org/10.1371/journal.pone.0068096
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author Mao, Junxia
Lv, Jia
Miao, Yan
Sun, Changsen
Hu, Liping
Zhang, Ru
Fu, Xiaoteng
Zhang, Lingling
Hu, Xiaoli
Wang, Shi
Bao, Zhenmin
author_facet Mao, Junxia
Lv, Jia
Miao, Yan
Sun, Changsen
Hu, Liping
Zhang, Ru
Fu, Xiaoteng
Zhang, Lingling
Hu, Xiaoli
Wang, Shi
Bao, Zhenmin
author_sort Mao, Junxia
collection PubMed
description Non-lethal DNA sampling has long appealed to researchers studying population and conservation genetics, as it does not necessitate removing individuals permanently from their natural environment or destroying valuable samples. However, such an approach has not yet been well established in bivalves. In this study, we demonstrate that the gill represents a good source of tissue for non-lethal sampling in scallops. Removal of a few gill filaments caused no noticeable behavioral abnormalities or increased mortality rates in Zhikong scallop (Chlamys farreri) during a three-month period of observation. To facilitate rapid gill-based DNA extraction, six methods (MA-MF) were designed and evaluated, each requiring less than one hour of processing time. The optimal method was identified as MF, in terms of maintaining DNA integrity and genotyping accuracy. Further optimization of MF method by orthogonal experimental design suggested that the utilization of gills could be limited to 2 mg of sample, which is sufficient for performing up to 20,000 PCR reactions. We also demonstrate the excellent cross-species utility of MF in two additional scallop species, Yesso scallop (Patinopecten yessoensis) and bay scallop (Argopecten irradians). Taken together, our study provides a rapid and efficient approach for applying non-lethal DNA sampling in bivalve species, which would serve as a valuable tool for maintaining bivalve populations and conservation genetics, as well as in breeding studies.
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spelling pubmed-37066022013-07-19 Development of a Rapid and Efficient Method for Non-Lethal DNA Sampling and Genotyping in Scallops Mao, Junxia Lv, Jia Miao, Yan Sun, Changsen Hu, Liping Zhang, Ru Fu, Xiaoteng Zhang, Lingling Hu, Xiaoli Wang, Shi Bao, Zhenmin PLoS One Research Article Non-lethal DNA sampling has long appealed to researchers studying population and conservation genetics, as it does not necessitate removing individuals permanently from their natural environment or destroying valuable samples. However, such an approach has not yet been well established in bivalves. In this study, we demonstrate that the gill represents a good source of tissue for non-lethal sampling in scallops. Removal of a few gill filaments caused no noticeable behavioral abnormalities or increased mortality rates in Zhikong scallop (Chlamys farreri) during a three-month period of observation. To facilitate rapid gill-based DNA extraction, six methods (MA-MF) were designed and evaluated, each requiring less than one hour of processing time. The optimal method was identified as MF, in terms of maintaining DNA integrity and genotyping accuracy. Further optimization of MF method by orthogonal experimental design suggested that the utilization of gills could be limited to 2 mg of sample, which is sufficient for performing up to 20,000 PCR reactions. We also demonstrate the excellent cross-species utility of MF in two additional scallop species, Yesso scallop (Patinopecten yessoensis) and bay scallop (Argopecten irradians). Taken together, our study provides a rapid and efficient approach for applying non-lethal DNA sampling in bivalve species, which would serve as a valuable tool for maintaining bivalve populations and conservation genetics, as well as in breeding studies. Public Library of Science 2013-07-09 /pmc/articles/PMC3706602/ /pubmed/23874509 http://dx.doi.org/10.1371/journal.pone.0068096 Text en © 2013 Mao et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Mao, Junxia
Lv, Jia
Miao, Yan
Sun, Changsen
Hu, Liping
Zhang, Ru
Fu, Xiaoteng
Zhang, Lingling
Hu, Xiaoli
Wang, Shi
Bao, Zhenmin
Development of a Rapid and Efficient Method for Non-Lethal DNA Sampling and Genotyping in Scallops
title Development of a Rapid and Efficient Method for Non-Lethal DNA Sampling and Genotyping in Scallops
title_full Development of a Rapid and Efficient Method for Non-Lethal DNA Sampling and Genotyping in Scallops
title_fullStr Development of a Rapid and Efficient Method for Non-Lethal DNA Sampling and Genotyping in Scallops
title_full_unstemmed Development of a Rapid and Efficient Method for Non-Lethal DNA Sampling and Genotyping in Scallops
title_short Development of a Rapid and Efficient Method for Non-Lethal DNA Sampling and Genotyping in Scallops
title_sort development of a rapid and efficient method for non-lethal dna sampling and genotyping in scallops
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3706602/
https://www.ncbi.nlm.nih.gov/pubmed/23874509
http://dx.doi.org/10.1371/journal.pone.0068096
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