Selection of reference genes for expression studies with fish myogenic cell cultures
Background Relatively few studies have used cell culture systems to investigate gene expression and the regulation of myogenesis in fish. To produce robust data from quantitative real-time PCR mRNA levels need to be normalised using internal reference genes which have stable expression across all ex...
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ftunivqespace:oai:espace.library.uq.edu.au:UQ:aa2e7bc 2023-05-15T15:31:24+02:00 Selection of reference genes for expression studies with fish myogenic cell cultures Bower, Neil I. Johnston, Ian A. William Hunter III 2011-01-01 https://espace.library.uq.edu.au/view/UQ:aa2e7bc eng eng Apple Academic Press doi:10.1201/b14534 orcid:0000-0002-6764-6063 1100 Agricultural and Biological Sciences 2200 Engineering Book Chapter 2011 ftunivqespace https://doi.org/10.1201/b14534 2020-08-06T13:45:53Z Background Relatively few studies have used cell culture systems to investigate gene expression and the regulation of myogenesis in fish. To produce robust data from quantitative real-time PCR mRNA levels need to be normalised using internal reference genes which have stable expression across all experimental samples. We have investigated the expression of eight candidate genes to identify suitable reference genes for use in primary myogenic cell cultures from Atlantic salmon (Salmo salar L.). The software analysis packages geNorm, Normfinder and Best keeper were used to rank genes according to their stability across 42 samples during the course of myogenic differentiation. Results Initial results showed several of the candidate genes exhibited stable expression throughout myogenic culture while Sdha was identified as the least stable gene. Further analysis with geNorm, Normfinder and Bestkeeper identified Ef1α, Hprt1, Ppia and RNApolII as stably expressed. Comparison of data normalised with the geometric average obtained from combinations of any three of these genes showed no significant differences, indicating that any combination of these genes is valid. Conclusion The geometric average of any three of Hprt1, Ef1α, Ppia and RNApolII is suitable for normalisation of gene expression data in primary myogenic cultures from Atlantic salmon. Book Part Atlantic salmon Salmo salar The University of Queensland: UQ eSpace |
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The University of Queensland: UQ eSpace |
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ftunivqespace |
language |
English |
topic |
1100 Agricultural and Biological Sciences 2200 Engineering |
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1100 Agricultural and Biological Sciences 2200 Engineering Bower, Neil I. Johnston, Ian A. Selection of reference genes for expression studies with fish myogenic cell cultures |
topic_facet |
1100 Agricultural and Biological Sciences 2200 Engineering |
description |
Background Relatively few studies have used cell culture systems to investigate gene expression and the regulation of myogenesis in fish. To produce robust data from quantitative real-time PCR mRNA levels need to be normalised using internal reference genes which have stable expression across all experimental samples. We have investigated the expression of eight candidate genes to identify suitable reference genes for use in primary myogenic cell cultures from Atlantic salmon (Salmo salar L.). The software analysis packages geNorm, Normfinder and Best keeper were used to rank genes according to their stability across 42 samples during the course of myogenic differentiation. Results Initial results showed several of the candidate genes exhibited stable expression throughout myogenic culture while Sdha was identified as the least stable gene. Further analysis with geNorm, Normfinder and Bestkeeper identified Ef1α, Hprt1, Ppia and RNApolII as stably expressed. Comparison of data normalised with the geometric average obtained from combinations of any three of these genes showed no significant differences, indicating that any combination of these genes is valid. Conclusion The geometric average of any three of Hprt1, Ef1α, Ppia and RNApolII is suitable for normalisation of gene expression data in primary myogenic cultures from Atlantic salmon. |
author2 |
William Hunter III |
format |
Book Part |
author |
Bower, Neil I. Johnston, Ian A. |
author_facet |
Bower, Neil I. Johnston, Ian A. |
author_sort |
Bower, Neil I. |
title |
Selection of reference genes for expression studies with fish myogenic cell cultures |
title_short |
Selection of reference genes for expression studies with fish myogenic cell cultures |
title_full |
Selection of reference genes for expression studies with fish myogenic cell cultures |
title_fullStr |
Selection of reference genes for expression studies with fish myogenic cell cultures |
title_full_unstemmed |
Selection of reference genes for expression studies with fish myogenic cell cultures |
title_sort |
selection of reference genes for expression studies with fish myogenic cell cultures |
publisher |
Apple Academic Press |
publishDate |
2011 |
url |
https://espace.library.uq.edu.au/view/UQ:aa2e7bc |
genre |
Atlantic salmon Salmo salar |
genre_facet |
Atlantic salmon Salmo salar |
op_relation |
doi:10.1201/b14534 orcid:0000-0002-6764-6063 |
op_doi |
https://doi.org/10.1201/b14534 |
_version_ |
1766361893605212160 |