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Comparison and optimization for DNA extraction of archived fish specimens

The DNA extracted from museum alcohol-fixed specimens can be a valuable source of information for solving taxonomic, phylogenetic, ecological and conservational questions. However, this type of DNA, also called ancient DNA, is routinely obtained in small portions and highly fragmented. We have teste...

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Autores principales: Silva, Priscilla C., Malabarba, Maria Claudia, Vari (In memoriam), Richard, Malabarba, Luiz R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6584492/
https://www.ncbi.nlm.nih.gov/pubmed/31249793
http://dx.doi.org/10.1016/j.mex.2019.06.001
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author Silva, Priscilla C.
Malabarba, Maria Claudia
Vari (In memoriam), Richard
Malabarba, Luiz R.
author_facet Silva, Priscilla C.
Malabarba, Maria Claudia
Vari (In memoriam), Richard
Malabarba, Luiz R.
author_sort Silva, Priscilla C.
collection PubMed
description The DNA extracted from museum alcohol-fixed specimens can be a valuable source of information for solving taxonomic, phylogenetic, ecological and conservational questions. However, this type of DNA, also called ancient DNA, is routinely obtained in small portions and highly fragmented. We have tested two different extraction kits in museum type-specimens of the fish family Characidae. Aiming to increase the DNA yield, we made modifications on a Qiagen manufacturer protocol, in the elution step. Also, to overcome the issue of DNA fragmentation, we applied our efforts in Sanger sequencing, to find a highly variable and, in result, informative COI fragment. Based on our results, there is no correlation between amount of the DNA extracted and the age of the sample. The Sanger sequencing generated sequences which are useful in solving taxonomic puzzles. Here are presented the customization and guidelines that allowed us to recover DNA from the archived fish specimens. • DNA extraction from archived fish specimens is more effective when using silica columns. • Change of the elution times from minutes in room temperature to 24 h in freezer greatly improved the DNA yielded. • Short but highly variable sequences replace the need to sequence the entire gene to identify a species.
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spelling pubmed-65844922019-06-27 Comparison and optimization for DNA extraction of archived fish specimens Silva, Priscilla C. Malabarba, Maria Claudia Vari (In memoriam), Richard Malabarba, Luiz R. MethodsX Biochemistry, Genetics and Molecular Biology The DNA extracted from museum alcohol-fixed specimens can be a valuable source of information for solving taxonomic, phylogenetic, ecological and conservational questions. However, this type of DNA, also called ancient DNA, is routinely obtained in small portions and highly fragmented. We have tested two different extraction kits in museum type-specimens of the fish family Characidae. Aiming to increase the DNA yield, we made modifications on a Qiagen manufacturer protocol, in the elution step. Also, to overcome the issue of DNA fragmentation, we applied our efforts in Sanger sequencing, to find a highly variable and, in result, informative COI fragment. Based on our results, there is no correlation between amount of the DNA extracted and the age of the sample. The Sanger sequencing generated sequences which are useful in solving taxonomic puzzles. Here are presented the customization and guidelines that allowed us to recover DNA from the archived fish specimens. • DNA extraction from archived fish specimens is more effective when using silica columns. • Change of the elution times from minutes in room temperature to 24 h in freezer greatly improved the DNA yielded. • Short but highly variable sequences replace the need to sequence the entire gene to identify a species. Elsevier 2019-06-08 /pmc/articles/PMC6584492/ /pubmed/31249793 http://dx.doi.org/10.1016/j.mex.2019.06.001 Text en © 2019 The Authors http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Biochemistry, Genetics and Molecular Biology
Silva, Priscilla C.
Malabarba, Maria Claudia
Vari (In memoriam), Richard
Malabarba, Luiz R.
Comparison and optimization for DNA extraction of archived fish specimens
title Comparison and optimization for DNA extraction of archived fish specimens
title_full Comparison and optimization for DNA extraction of archived fish specimens
title_fullStr Comparison and optimization for DNA extraction of archived fish specimens
title_full_unstemmed Comparison and optimization for DNA extraction of archived fish specimens
title_short Comparison and optimization for DNA extraction of archived fish specimens
title_sort comparison and optimization for dna extraction of archived fish specimens
topic Biochemistry, Genetics and Molecular Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6584492/
https://www.ncbi.nlm.nih.gov/pubmed/31249793
http://dx.doi.org/10.1016/j.mex.2019.06.001
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