Abstract

To better understand host and immune response to diseases, gene expression studies require identification of reference genes with stable expression for accurate normalisation. This study describes the identification and testing of reference genes with stable expression profiles in koala lymph node tissues across two genetically distinct koala populations. From the 25 most stable genes identified in transcriptome analysis, 11 genes were selected for verification using reverse transcription quantitative PCR, in addition to the commonly used ACTB and GAPDH genes. The expression data were analysed using stable genes statistical software - geNorm, BestKeeper, NormFinder, the comparative ΔCt method and RefFinder. All 13 genes showed relative stability in expression in koala lymph node tissues, however Tmem97 and Hmg20a were identified as the most stable genes across the two koala populations.

Details

Title
Identification of stable reference genes for quantitative PCR in koalas
Author
Sarker, N 1 ; Fabijan, J 2 ; Emes, R D 3 ; Hemmatzadeh, F 2 ; Meers, J 1 ; Moreton, J 4 ; Owen, H 1 ; Seddon, J M 1 ; Simmons, G 1 ; Speight, N 2 ; Trott, D 2 ; Woolford, L 2 ; Tarlinton, R E 5   VIAFID ORCID Logo 

1  School of Veterinary Science, The University of Queensland, Gatton, QLD, Australia 
2  School of Animal and Veterinary Sciences, The University of Adelaide, Roseworthy, SA, Australia 
3  School of Veterinary Medicine and Science, University of Nottingham, Sutton Bonington Campus, Loughborough, United Kingdom; Advanced Data Analysis Centre (ADAC), University of Nottingham, Sutton Bonington, United Kingdom 
4  Advanced Data Analysis Centre (ADAC), University of Nottingham, Sutton Bonington, United Kingdom 
5  School of Veterinary Medicine and Science, University of Nottingham, Sutton Bonington Campus, Loughborough, United Kingdom 
Pages
1-8
Publication year
2018
Publication date
Feb 2018
Publisher
Nature Publishing Group
e-ISSN
20452322
Source type
Scholarly Journal
Language of publication
English
DOI
https://doi.org/10.1038/s41598-018-21723-0
ProQuest document ID
2006814574
Copyright
© 2018. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.