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Identification of stable reference genes for quantitative PCR in koalas

Sarker, N.; Fabijan, J.; Emes, Richard D.; Hemmatzadeh, F.; Meers, J.; Moreton, Joanna; Owen, H.; Seddon, J.; Simmons, G.; Speight, N.; Trott, D.; Woolford, L.; Tarlinton, Rachael E.

Authors

N. Sarker

J. Fabijan

RICHARD EMES richard.emes@nottingham.ac.uk
Professor of Bioinformatics

F. Hemmatzadeh

J. Meers

Joanna Moreton

H. Owen

J. Seddon

G. Simmons

N. Speight

D. Trott

L. Woolford



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.

Citation

Sarker, N., Fabijan, J., Emes, R. D., Hemmatzadeh, F., Meers, J., Moreton, J., …Tarlinton, R. E. (in press). Identification of stable reference genes for quantitative PCR in koalas. Scientific Reports, 8, https://doi.org/10.1038/s41598-018-21723-0

Journal Article Type Article
Acceptance Date Feb 9, 2018
Online Publication Date Feb 20, 2018
Deposit Date Feb 23, 2018
Publicly Available Date Feb 23, 2018
Journal Scientific Reports
Print ISSN 2045-2322
Electronic ISSN 2045-2322
Publisher Nature Publishing Group
Peer Reviewed Peer Reviewed
Volume 8
Article Number 3364
DOI https://doi.org/10.1038/s41598-018-21723-0
Keywords stable gene, koala, RT-qPCR
Public URL http://eprints.nottingham.ac.uk/id/eprint/49705
Publisher URL https://www.nature.com/articles/s41598-018-21723-0
Copyright Statement Copyright information regarding this work can be found at the following address: http://creativecommons.org/licenses/by/4.0

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Copyright Statement
Copyright information regarding this work can be found at the following address: http://creativecommons.org/licenses/by/4.0


s41598-018-21723-0.pdf (1.5 Mb)
PDF

Copyright Statement
Copyright information regarding this work can be found at the following address: http://creativecommons.org/licenses/by/4.0





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