A comparison of normalization methods for the expression of genes associated with oxidative stress in the liver of sheep.

Sarah Babington, Luoyang Ding, Alan J Tilbrook, Shane K Maloney, Dominique Blache

Abstract readComparative Study

In one paragraph

Article in BMC genomic data, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 1 paper.

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1citing papers in PubMed

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1 · What the graph read from it

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1 citing paper in PubMed.

Genomic Signatures of Selection Are Enriched in Differentially Expressed Genes in Sticklebacks Adapting to Contrasting Environments.Genome biology and evolution · 2026
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5 · Who and what money

Authors and funding

5 authors.

Sarah BabingtonSchool of Agriculture and Environment, The University of Western Australia, Crawley, WA, 6009, Australia.

Luoyang DingSchool of Agriculture and Environment, The University of Western Australia, Crawley, WA, 6009, Australia.

Alan J TilbrookSchool of Veterinary Science, The University of Queensland, Gatton, QLD, 4343, Australia.

Shane K MaloneySchool of Human Sciences, The University of Western Australia, Crawley, WA, 6009, Australia.

Dominique BlacheSchool of Agriculture and Environment, The University of Western Australia, Crawley, WA, 6009, Australia. dominique.blache@uwa.edu.au.

Funding

Meat and Livestock Australia P.PSH.1232

6 · The paper itself

Abstract

backgroundNormalization of data obtained from RT-qPCR studies is important for accurate interpretation of the results. The two most common methods of normalization that are used are the reference gene method or an algorithm-only approach, such as NORMA-Gene. Here, we assessed the impact of normalization using reference genes or the NORMA-Gene method on the expression results of five target genes that are related to oxidative stress (CAT, GPX1, GPX3, PRDX1, and SOD1) in the liver of sheep that had been exposed to three dietary treatments.

resultsThe reference genes selected as the most stable across samples for normalization were HPRT1, HSP90AA1, and B2M. Interpretation of the effect of the treatment on the expression of GPX3 differed significantly between the methods of normalization. NORMA-Gene was better at reducing the variance in the expression of the target genes than was any of the other normalization methods.

conclusionsWe demonstrated that NORMA-Gene can provide a more reliable normalization method that requires less resources than the use of reference genes for studies on gene expression in livestock.

Indexed as

Gene Expression ProfilingLiverOxidative StressAnimalsGlutathione PeroxidaseReal-Time Polymerase Chain ReactionReference StandardsSheepGlutathione PeroxidaseGene expressionNORMA-GeneOvineReference genesRT-qPCR

Identifiers

PMID40745265

DOI10.1186/s12863-025-01345-y

PMCPMC12312509

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LicenceCC BY-NC-ND

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