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High-Resolution Melting of 12S rRNA and Cytochrome b DNA Sequences for Discrimination of Species within Distinct European Animal Families
The cheap and easy identification of species is necessary within multiple fields of molecular biology. The use of high-resolution melting (HRM) of DNA provides a fast closed-tube method for analysis of the sequence composition of the mitochondrial genes 12S rRNA and cytochrome b. We investigated the...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4274031/ https://www.ncbi.nlm.nih.gov/pubmed/25531439 http://dx.doi.org/10.1371/journal.pone.0115575 |
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author | Naue, Jana Hansmann, Tobias Schmidt, Ulrike |
author_facet | Naue, Jana Hansmann, Tobias Schmidt, Ulrike |
author_sort | Naue, Jana |
collection | PubMed |
description | The cheap and easy identification of species is necessary within multiple fields of molecular biology. The use of high-resolution melting (HRM) of DNA provides a fast closed-tube method for analysis of the sequence composition of the mitochondrial genes 12S rRNA and cytochrome b. We investigated the potential use of HRM for species identification within eleven different animal groups commonly found in Europe by animal-group-specific DNA amplification followed by DNA melting. Influence factors as DNA amount, additional single base alterations, and the existence of mixed samples were taken into consideration. Visual inspection combined with mathematical evaluation of the curve shapes did resolve nearly all species within an animal group. The assay can therefore not only be used for identification of animal groups and mixture analysis but also for species identification within the respective groups. The use of a universal 12S rRNA system additionally revealed a possible approach for species discrimination, mostly by exclusion. The use of the HRM assay showed to be a reliable, fast, and cheap method for species discrimination within a broad range of different animal species and can be used in a flexible “modular” manner depending on the question to be solved. |
format | Online Article Text |
id | pubmed-4274031 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-42740312014-12-31 High-Resolution Melting of 12S rRNA and Cytochrome b DNA Sequences for Discrimination of Species within Distinct European Animal Families Naue, Jana Hansmann, Tobias Schmidt, Ulrike PLoS One Research Article The cheap and easy identification of species is necessary within multiple fields of molecular biology. The use of high-resolution melting (HRM) of DNA provides a fast closed-tube method for analysis of the sequence composition of the mitochondrial genes 12S rRNA and cytochrome b. We investigated the potential use of HRM for species identification within eleven different animal groups commonly found in Europe by animal-group-specific DNA amplification followed by DNA melting. Influence factors as DNA amount, additional single base alterations, and the existence of mixed samples were taken into consideration. Visual inspection combined with mathematical evaluation of the curve shapes did resolve nearly all species within an animal group. The assay can therefore not only be used for identification of animal groups and mixture analysis but also for species identification within the respective groups. The use of a universal 12S rRNA system additionally revealed a possible approach for species discrimination, mostly by exclusion. The use of the HRM assay showed to be a reliable, fast, and cheap method for species discrimination within a broad range of different animal species and can be used in a flexible “modular” manner depending on the question to be solved. Public Library of Science 2014-12-22 /pmc/articles/PMC4274031/ /pubmed/25531439 http://dx.doi.org/10.1371/journal.pone.0115575 Text en © 2014 Naue 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 Naue, Jana Hansmann, Tobias Schmidt, Ulrike High-Resolution Melting of 12S rRNA and Cytochrome b DNA Sequences for Discrimination of Species within Distinct European Animal Families |
title | High-Resolution Melting of 12S rRNA and Cytochrome b DNA Sequences for Discrimination of Species within Distinct European Animal Families |
title_full | High-Resolution Melting of 12S rRNA and Cytochrome b DNA Sequences for Discrimination of Species within Distinct European Animal Families |
title_fullStr | High-Resolution Melting of 12S rRNA and Cytochrome b DNA Sequences for Discrimination of Species within Distinct European Animal Families |
title_full_unstemmed | High-Resolution Melting of 12S rRNA and Cytochrome b DNA Sequences for Discrimination of Species within Distinct European Animal Families |
title_short | High-Resolution Melting of 12S rRNA and Cytochrome b DNA Sequences for Discrimination of Species within Distinct European Animal Families |
title_sort | high-resolution melting of 12s rrna and cytochrome b dna sequences for discrimination of species within distinct european animal families |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4274031/ https://www.ncbi.nlm.nih.gov/pubmed/25531439 http://dx.doi.org/10.1371/journal.pone.0115575 |
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