Evaluation of potential reference genes in real-time RT-PCR studies of Atlantic salmon
Background Salmonid fishes are among the most widely studied model fish species but reports on systematic evaluation of reference genes in qRT-PCR studies is lacking. Results The stability of six potential reference genes was examined in eight tissues of Atlantic salmon (Salmo salar), to determine t...
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Online Access: | http://hdl.handle.net/11250/2636174 https://doi.org/10.1186/1471-2199-6-21 |
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ftimr:oai:imr.brage.unit.no:11250/2636174 2023-05-15T15:30:08+02:00 Evaluation of potential reference genes in real-time RT-PCR studies of Atlantic salmon Olsvik, Pål A. Lie, Kai Kristoffer Olderbakk Jordal, Anne-Lise Nilsen, Tom Ole Hordvik, Ivar 2005 application/pdf http://hdl.handle.net/11250/2636174 https://doi.org/10.1186/1471-2199-6-21 eng eng BMC Molecular Biology. 2005, 6 (21), 1-9. urn:issn:1471-2199 http://hdl.handle.net/11250/2636174 https://doi.org/10.1186/1471-2199-6-21 cristin:364648 1-9 6 BMC Molecular Biology 21 Journal article Peer reviewed 2005 ftimr https://doi.org/10.1186/1471-2199-6-21 2021-09-23T20:15:57Z Background Salmonid fishes are among the most widely studied model fish species but reports on systematic evaluation of reference genes in qRT-PCR studies is lacking. Results The stability of six potential reference genes was examined in eight tissues of Atlantic salmon (Salmo salar), to determine the most suitable genes to be used in quantitative real-time RT-PCR analyses. The relative transcription levels of genes encoding 18S rRNA, S20 ribosomal protein, β-actin, glyceraldehyde-3P-dehydrogenase (GAPDH), and two paralog genes encoding elongation factor 1A (EF1AA and EF1AB) were quantified in gills, liver, head kidney, spleen, thymus, brain, muscle, and posterior intestine in six untreated adult fish, in addition to a group of individuals that went through smoltification. Based on calculations performed with the geNorm VBA applet, which determines the most stable genes from a set of tested genes in a given cDNA sample, the ranking of the examined genes in adult Atlantic salmon was EF1AB>EF1AA>β-actin>18S rRNA>S20>GAPDH. When the same calculations were done on a total of 24 individuals from four stages in the smoltification process (presmolt, smolt, smoltified seawater and desmoltified freshwater), the gene ranking was EF1AB>EF1AA>S20>β-actin>18S rRNA>GAPDH. Conclusion Overall, this work suggests that the EF1AA and EF1AB genes can be useful as reference genes in qRT-PCR examination of gene expression in the Atlantic salmon. publishedVersion Article in Journal/Newspaper Atlantic salmon Salmo salar Institute for Marine Research: Brage IMR BMC Molecular Biology 6 1 21 |
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Institute for Marine Research: Brage IMR |
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language |
English |
description |
Background Salmonid fishes are among the most widely studied model fish species but reports on systematic evaluation of reference genes in qRT-PCR studies is lacking. Results The stability of six potential reference genes was examined in eight tissues of Atlantic salmon (Salmo salar), to determine the most suitable genes to be used in quantitative real-time RT-PCR analyses. The relative transcription levels of genes encoding 18S rRNA, S20 ribosomal protein, β-actin, glyceraldehyde-3P-dehydrogenase (GAPDH), and two paralog genes encoding elongation factor 1A (EF1AA and EF1AB) were quantified in gills, liver, head kidney, spleen, thymus, brain, muscle, and posterior intestine in six untreated adult fish, in addition to a group of individuals that went through smoltification. Based on calculations performed with the geNorm VBA applet, which determines the most stable genes from a set of tested genes in a given cDNA sample, the ranking of the examined genes in adult Atlantic salmon was EF1AB>EF1AA>β-actin>18S rRNA>S20>GAPDH. When the same calculations were done on a total of 24 individuals from four stages in the smoltification process (presmolt, smolt, smoltified seawater and desmoltified freshwater), the gene ranking was EF1AB>EF1AA>S20>β-actin>18S rRNA>GAPDH. Conclusion Overall, this work suggests that the EF1AA and EF1AB genes can be useful as reference genes in qRT-PCR examination of gene expression in the Atlantic salmon. publishedVersion |
format |
Article in Journal/Newspaper |
author |
Olsvik, Pål A. Lie, Kai Kristoffer Olderbakk Jordal, Anne-Lise Nilsen, Tom Ole Hordvik, Ivar |
spellingShingle |
Olsvik, Pål A. Lie, Kai Kristoffer Olderbakk Jordal, Anne-Lise Nilsen, Tom Ole Hordvik, Ivar Evaluation of potential reference genes in real-time RT-PCR studies of Atlantic salmon |
author_facet |
Olsvik, Pål A. Lie, Kai Kristoffer Olderbakk Jordal, Anne-Lise Nilsen, Tom Ole Hordvik, Ivar |
author_sort |
Olsvik, Pål A. |
title |
Evaluation of potential reference genes in real-time RT-PCR studies of Atlantic salmon |
title_short |
Evaluation of potential reference genes in real-time RT-PCR studies of Atlantic salmon |
title_full |
Evaluation of potential reference genes in real-time RT-PCR studies of Atlantic salmon |
title_fullStr |
Evaluation of potential reference genes in real-time RT-PCR studies of Atlantic salmon |
title_full_unstemmed |
Evaluation of potential reference genes in real-time RT-PCR studies of Atlantic salmon |
title_sort |
evaluation of potential reference genes in real-time rt-pcr studies of atlantic salmon |
publishDate |
2005 |
url |
http://hdl.handle.net/11250/2636174 https://doi.org/10.1186/1471-2199-6-21 |
genre |
Atlantic salmon Salmo salar |
genre_facet |
Atlantic salmon Salmo salar |
op_source |
1-9 6 BMC Molecular Biology 21 |
op_relation |
BMC Molecular Biology. 2005, 6 (21), 1-9. urn:issn:1471-2199 http://hdl.handle.net/11250/2636174 https://doi.org/10.1186/1471-2199-6-21 cristin:364648 |
op_doi |
https://doi.org/10.1186/1471-2199-6-21 |
container_title |
BMC Molecular Biology |
container_volume |
6 |
container_issue |
1 |
container_start_page |
21 |
_version_ |
1766360575829344256 |