Preservation of RNA and DNA from mammal samples under field conditions
Ecological and conservation genetics require sampling of organisms in the wild. Appropriate preservation of the collected samples, usually by cryostorage, is key to the quality of the genetic data obtained. Nevertheless, cryopreservation in the field to ensure RNA and DNA stability is not always pos...
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2013
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Online Access: | http://hdl.handle.net/10261/80402 https://doi.org/10.1111/1755-0998.12108 |
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ftcsic:oai:digital.csic.es:10261/80402 2024-02-11T10:08:13+01:00 Preservation of RNA and DNA from mammal samples under field conditions Camacho-Sánchez, Miguel Burraco, Pablo Gómez-Mestre, Iván Leonard, Jennifer A. 2013 http://hdl.handle.net/10261/80402 https://doi.org/10.1111/1755-0998.12108 en eng Blackwell Publishing doi:10.1111/1755-0998.12108 issn: 1755-098X Molecular Ecology Resources 13: 663- 673 (2013) http://hdl.handle.net/10261/80402 open artículo http://purl.org/coar/resource_type/c_6501 2013 ftcsic https://doi.org/10.1111/1755-0998.12108 2024-01-16T09:51:17Z Ecological and conservation genetics require sampling of organisms in the wild. Appropriate preservation of the collected samples, usually by cryostorage, is key to the quality of the genetic data obtained. Nevertheless, cryopreservation in the field to ensure RNA and DNA stability is not always possible. We compared several nucleic acid preservation solutions appropriate for field sampling and tested them on rat (Rattus rattus) blood, ear and tail tip, liver, brain and muscle. We compared the efficacy of a nucleic acid preservation (NAP) buffer for DNA preservation against 95% ethanol and Longmire buffer, and for RNA preservation against RNAlater (Qiagen) and Longmire buffer, under simulated field conditions. For DNA, the NAP buffer was slightly better than cryopreservation or 95% ethanol, but high molecular weight DNA was preserved in all conditions. The NAP buffer preserved RNA as well as RNAlater. Liver yielded the best RNA and DNA quantity and quality; thus, liver should be the tissue preferentially collected from euthanized animals. We also show that DNA persists in nonpreserved muscle tissue for at least 1 week at ambient temperature, although degradation is noticeable in a matter of hours. When cryopreservation is not possible, the NAP buffer is an economical alternative for RNA preservation at ambient temperature for at least 2 months and DNA preservation for at least 10 months. © 2013 John Wiley & Sons Ltd. Peer Reviewed Article in Journal/Newspaper Rattus rattus Digital.CSIC (Spanish National Research Council) Molecular Ecology Resources 13 4 663 673 |
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Digital.CSIC (Spanish National Research Council) |
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English |
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Ecological and conservation genetics require sampling of organisms in the wild. Appropriate preservation of the collected samples, usually by cryostorage, is key to the quality of the genetic data obtained. Nevertheless, cryopreservation in the field to ensure RNA and DNA stability is not always possible. We compared several nucleic acid preservation solutions appropriate for field sampling and tested them on rat (Rattus rattus) blood, ear and tail tip, liver, brain and muscle. We compared the efficacy of a nucleic acid preservation (NAP) buffer for DNA preservation against 95% ethanol and Longmire buffer, and for RNA preservation against RNAlater (Qiagen) and Longmire buffer, under simulated field conditions. For DNA, the NAP buffer was slightly better than cryopreservation or 95% ethanol, but high molecular weight DNA was preserved in all conditions. The NAP buffer preserved RNA as well as RNAlater. Liver yielded the best RNA and DNA quantity and quality; thus, liver should be the tissue preferentially collected from euthanized animals. We also show that DNA persists in nonpreserved muscle tissue for at least 1 week at ambient temperature, although degradation is noticeable in a matter of hours. When cryopreservation is not possible, the NAP buffer is an economical alternative for RNA preservation at ambient temperature for at least 2 months and DNA preservation for at least 10 months. © 2013 John Wiley & Sons Ltd. Peer Reviewed |
format |
Article in Journal/Newspaper |
author |
Camacho-Sánchez, Miguel Burraco, Pablo Gómez-Mestre, Iván Leonard, Jennifer A. |
spellingShingle |
Camacho-Sánchez, Miguel Burraco, Pablo Gómez-Mestre, Iván Leonard, Jennifer A. Preservation of RNA and DNA from mammal samples under field conditions |
author_facet |
Camacho-Sánchez, Miguel Burraco, Pablo Gómez-Mestre, Iván Leonard, Jennifer A. |
author_sort |
Camacho-Sánchez, Miguel |
title |
Preservation of RNA and DNA from mammal samples under field conditions |
title_short |
Preservation of RNA and DNA from mammal samples under field conditions |
title_full |
Preservation of RNA and DNA from mammal samples under field conditions |
title_fullStr |
Preservation of RNA and DNA from mammal samples under field conditions |
title_full_unstemmed |
Preservation of RNA and DNA from mammal samples under field conditions |
title_sort |
preservation of rna and dna from mammal samples under field conditions |
publisher |
Blackwell Publishing |
publishDate |
2013 |
url |
http://hdl.handle.net/10261/80402 https://doi.org/10.1111/1755-0998.12108 |
genre |
Rattus rattus |
genre_facet |
Rattus rattus |
op_relation |
doi:10.1111/1755-0998.12108 issn: 1755-098X Molecular Ecology Resources 13: 663- 673 (2013) http://hdl.handle.net/10261/80402 |
op_rights |
open |
op_doi |
https://doi.org/10.1111/1755-0998.12108 |
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Molecular Ecology Resources |
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13 |
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4 |
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663 |
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673 |
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1790607245779664896 |