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Combined hybridization capture and shotgun sequencing for ancient DNA analysis of extinct wild and domestic dromedary camel
The performance of hybridization capture combined with next‐generation sequencing (NGS) has seen limited investigation with samples from hot and arid regions until now. We applied hybridization capture and shotgun sequencing to recover DNA sequences from bone specimens of ancient‐domestic dromedary...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5324683/ https://www.ncbi.nlm.nih.gov/pubmed/27289015 http://dx.doi.org/10.1111/1755-0998.12551 |
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author | Mohandesan, Elmira Speller, Camilla F. Peters, Joris Uerpmann, Hans‐Peter Uerpmann, Margarethe De Cupere, Bea Hofreiter, Michael Burger, Pamela A. |
author_facet | Mohandesan, Elmira Speller, Camilla F. Peters, Joris Uerpmann, Hans‐Peter Uerpmann, Margarethe De Cupere, Bea Hofreiter, Michael Burger, Pamela A. |
author_sort | Mohandesan, Elmira |
collection | PubMed |
description | The performance of hybridization capture combined with next‐generation sequencing (NGS) has seen limited investigation with samples from hot and arid regions until now. We applied hybridization capture and shotgun sequencing to recover DNA sequences from bone specimens of ancient‐domestic dromedary (Camelus dromedarius) and its extinct ancestor, the wild dromedary from Jordan, Syria, Turkey and the Arabian Peninsula, respectively. Our results show that hybridization capture increased the percentage of mitochondrial DNA (mtDNA) recovery by an average 187‐fold and in some cases yielded virtually complete mitochondrial (mt) genomes at multifold coverage in a single capture experiment. Furthermore, we tested the effect of hybridization temperature and time by using a touchdown approach on a limited number of samples. We observed no significant difference in the number of unique dromedary mtDNA reads retrieved with the standard capture compared to the touchdown method. In total, we obtained 14 partial mitochondrial genomes from ancient‐domestic dromedaries with 17–95% length coverage and 1.27–47.1‐fold read depths for the covered regions. Using whole‐genome shotgun sequencing, we successfully recovered endogenous dromedary nuclear DNA (nuDNA) from domestic and wild dromedary specimens with 1–1.06‐fold read depths for covered regions. Our results highlight that despite recent methodological advances, obtaining ancient DNA (aDNA) from specimens recovered from hot, arid environments is still problematic. Hybridization protocols require specific optimization, and samples at the limit of DNA preservation need multiple replications of DNA extraction and hybridization capture as has been shown previously for Middle Pleistocene specimens. |
format | Online Article Text |
id | pubmed-5324683 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-53246832017-03-14 Combined hybridization capture and shotgun sequencing for ancient DNA analysis of extinct wild and domestic dromedary camel Mohandesan, Elmira Speller, Camilla F. Peters, Joris Uerpmann, Hans‐Peter Uerpmann, Margarethe De Cupere, Bea Hofreiter, Michael Burger, Pamela A. Mol Ecol Resour RESOURCE ARTICLES The performance of hybridization capture combined with next‐generation sequencing (NGS) has seen limited investigation with samples from hot and arid regions until now. We applied hybridization capture and shotgun sequencing to recover DNA sequences from bone specimens of ancient‐domestic dromedary (Camelus dromedarius) and its extinct ancestor, the wild dromedary from Jordan, Syria, Turkey and the Arabian Peninsula, respectively. Our results show that hybridization capture increased the percentage of mitochondrial DNA (mtDNA) recovery by an average 187‐fold and in some cases yielded virtually complete mitochondrial (mt) genomes at multifold coverage in a single capture experiment. Furthermore, we tested the effect of hybridization temperature and time by using a touchdown approach on a limited number of samples. We observed no significant difference in the number of unique dromedary mtDNA reads retrieved with the standard capture compared to the touchdown method. In total, we obtained 14 partial mitochondrial genomes from ancient‐domestic dromedaries with 17–95% length coverage and 1.27–47.1‐fold read depths for the covered regions. Using whole‐genome shotgun sequencing, we successfully recovered endogenous dromedary nuclear DNA (nuDNA) from domestic and wild dromedary specimens with 1–1.06‐fold read depths for covered regions. Our results highlight that despite recent methodological advances, obtaining ancient DNA (aDNA) from specimens recovered from hot, arid environments is still problematic. Hybridization protocols require specific optimization, and samples at the limit of DNA preservation need multiple replications of DNA extraction and hybridization capture as has been shown previously for Middle Pleistocene specimens. John Wiley and Sons Inc. 2016-08-01 2017-03 /pmc/articles/PMC5324683/ /pubmed/27289015 http://dx.doi.org/10.1111/1755-0998.12551 Text en © 2016 The Authors. Molecular Ecology Resources Published by John Wiley & Sons Ltd. This is an open access article under the terms of the Creative Commons Attribution (http://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | RESOURCE ARTICLES Mohandesan, Elmira Speller, Camilla F. Peters, Joris Uerpmann, Hans‐Peter Uerpmann, Margarethe De Cupere, Bea Hofreiter, Michael Burger, Pamela A. Combined hybridization capture and shotgun sequencing for ancient DNA analysis of extinct wild and domestic dromedary camel |
title | Combined hybridization capture and shotgun sequencing for ancient DNA analysis of extinct wild and domestic dromedary camel |
title_full | Combined hybridization capture and shotgun sequencing for ancient DNA analysis of extinct wild and domestic dromedary camel |
title_fullStr | Combined hybridization capture and shotgun sequencing for ancient DNA analysis of extinct wild and domestic dromedary camel |
title_full_unstemmed | Combined hybridization capture and shotgun sequencing for ancient DNA analysis of extinct wild and domestic dromedary camel |
title_short | Combined hybridization capture and shotgun sequencing for ancient DNA analysis of extinct wild and domestic dromedary camel |
title_sort | combined hybridization capture and shotgun sequencing for ancient dna analysis of extinct wild and domestic dromedary camel |
topic | RESOURCE ARTICLES |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5324683/ https://www.ncbi.nlm.nih.gov/pubmed/27289015 http://dx.doi.org/10.1111/1755-0998.12551 |
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