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A simple and efficient total genomic DNA extraction method for individual zooplankton

Molecular approaches are widely applied in species identification and taxonomic studies of minute zooplankton. One of the most focused zooplankton nowadays is from Subclass Copepoda. Accurate species identification of all life stages of the generally small sized copepods through molecular analysis i...

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Detalles Bibliográficos
Autores principales: Fazhan, Hanafiah, Waiho, Khor, Shahreza, Md. Sheriff
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer International Publishing 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5130931/
https://www.ncbi.nlm.nih.gov/pubmed/27995026
http://dx.doi.org/10.1186/s40064-016-3724-x
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author Fazhan, Hanafiah
Waiho, Khor
Shahreza, Md. Sheriff
author_facet Fazhan, Hanafiah
Waiho, Khor
Shahreza, Md. Sheriff
author_sort Fazhan, Hanafiah
collection PubMed
description Molecular approaches are widely applied in species identification and taxonomic studies of minute zooplankton. One of the most focused zooplankton nowadays is from Subclass Copepoda. Accurate species identification of all life stages of the generally small sized copepods through molecular analysis is important, especially in taxonomic and systematic assessment of harpacticoid copepod populations and to understand their dynamics within the marine community. However, total genomic DNA (TGDNA) extraction from individual harpacticoid copepods can be problematic due to their small size and epibenthic behavior. In this research, six TGDNA extraction methods done on individual harpacticoid copepods were compared. The first new simple, feasible, efficient and consistent TGDNA extraction method was designed and compared with the commercial kit and modified available TGDNA extraction methods. The newly described TGDNA extraction method, “Incubation in PCR buffer” method, yielded good and consistent results based on the high success rate of PCR amplification (82%) compared to other methods. Coupled with its relatively consistent and economical method the “Incubation in PCR buffer” method is highly recommended in the TGDNA extraction of other minute zooplankton species.
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spelling pubmed-51309312016-12-19 A simple and efficient total genomic DNA extraction method for individual zooplankton Fazhan, Hanafiah Waiho, Khor Shahreza, Md. Sheriff Springerplus Research Molecular approaches are widely applied in species identification and taxonomic studies of minute zooplankton. One of the most focused zooplankton nowadays is from Subclass Copepoda. Accurate species identification of all life stages of the generally small sized copepods through molecular analysis is important, especially in taxonomic and systematic assessment of harpacticoid copepod populations and to understand their dynamics within the marine community. However, total genomic DNA (TGDNA) extraction from individual harpacticoid copepods can be problematic due to their small size and epibenthic behavior. In this research, six TGDNA extraction methods done on individual harpacticoid copepods were compared. The first new simple, feasible, efficient and consistent TGDNA extraction method was designed and compared with the commercial kit and modified available TGDNA extraction methods. The newly described TGDNA extraction method, “Incubation in PCR buffer” method, yielded good and consistent results based on the high success rate of PCR amplification (82%) compared to other methods. Coupled with its relatively consistent and economical method the “Incubation in PCR buffer” method is highly recommended in the TGDNA extraction of other minute zooplankton species. Springer International Publishing 2016-12-01 /pmc/articles/PMC5130931/ /pubmed/27995026 http://dx.doi.org/10.1186/s40064-016-3724-x 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.
spellingShingle Research
Fazhan, Hanafiah
Waiho, Khor
Shahreza, Md. Sheriff
A simple and efficient total genomic DNA extraction method for individual zooplankton
title A simple and efficient total genomic DNA extraction method for individual zooplankton
title_full A simple and efficient total genomic DNA extraction method for individual zooplankton
title_fullStr A simple and efficient total genomic DNA extraction method for individual zooplankton
title_full_unstemmed A simple and efficient total genomic DNA extraction method for individual zooplankton
title_short A simple and efficient total genomic DNA extraction method for individual zooplankton
title_sort simple and efficient total genomic dna extraction method for individual zooplankton
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5130931/
https://www.ncbi.nlm.nih.gov/pubmed/27995026
http://dx.doi.org/10.1186/s40064-016-3724-x
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