Skip to content

Research at St Andrews

Selection of reference genes for expression studies with fish myogenic cell cultures

Research output: Contribution to journalArticle

Abstract

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 Ef1 alpha, 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, Ef1 a, Ppia and RNApolII is suitable for normalisation of gene expression data in primary myogenic cultures from Atlantic salmon.

Close

Details

Original languageEnglish
Article number80
Pages (from-to)-
Number of pages11
JournalBMC Molecular Biology
Volume10
DOIs
Publication statusPublished - 10 Aug 2009

    Research areas

  • Real-time PCR, Differential expression, Housekeeping genes, Skeletal-muscle, Myoblast fusion, Beta-actin, In-vitro, RT-PCR, Messenger, RNA

Discover related content
Find related publications, people, projects and more using interactive charts.

View graph of relations

Related by author

  1. Influence of feed ration size on somatic and muscle growth in landlocked dwarf and farmed Atlantic salmon Salmo salar

    Andersen, Ø., Vieira, V., Dessen, J-E. & Johnston, I. A., Apr 2019, In : Journal of Fish Biology. 94, 4, p. 614-620 7 p.

    Research output: Contribution to journalArticle

  2. Genomic tools and selective breeding in molluscs

    Hollenbeck, C. M. & Johnston, I. A., 18 Jul 2018, In : Frontiers in Genetics. 9, 15 p., 253.

    Research output: Contribution to journalReview article

  3. A collaborative European approach to accelerating translational marine science

    Brennecke, P., Ferrante, M., Johnston, I. A. & Smith, D., 5 Jul 2018, In : Journal of Marine Science and Engineering. 6, 3, 12 p., 81.

    Research output: Contribution to journalArticle

  4. A workflow used to design low density SNP panels for parentage assignment and traceability in aquaculture species and its validation in Atlantic salmon

    Holman, L. E., Garcia de la Serrana, D., Onoufriou, A., Hillestad, B. & Johnston, I. A., 1 Jul 2017, In : Aquaculture. 476, p. 59-64 6 p.

    Research output: Contribution to journalArticle

Related by journal

  1. BMC Molecular Biology (Journal)

    Stuart MacNeill (Editor)
    2011 → …

    Activity: Publication peer-review and editorial work typesEditor of research journal

  2. BMC Molecular Biology (Journal)

    Stuart MacNeill (Editor)
    20042011

    Activity: Publication peer-review and editorial work typesEditor of research journal

Related by journal

ID: 15933941