Babington Sarah, Ding Luoyang, Tilbrook Alan J, Maloney Shane K, Blache Dominique
School of Agriculture and Environment, The University of Western Australia, Crawley, WA, 6009, Australia.
School of Veterinary Science, The University of Queensland, Gatton, QLD, 4343, Australia.
BMC Genom Data. 2025 Jul 31;26(1):53. doi: 10.1186/s12863-025-01345-y.
Normalization 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 (, , ,, and ) in the liver of sheep that had been exposed to three dietary treatments.
The reference genes selected as the most stable across samples for normalization were , , and . Interpretation of the effect of the treatment on the expression of 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.
We 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.
The online version contains supplementary material available at 10.1186/s12863-025-01345-y.
对RT-qPCR研究获得的数据进行标准化处理对于准确解读结果至关重要。最常用的两种标准化方法是参考基因法或仅使用算法的方法，如NORMA-Gene。在此，我们评估了使用参考基因或NORMA-Gene方法进行标准化处理对暴露于三种饮食处理的绵羊肝脏中与氧化应激相关的五个靶基因（ 、 、 、 和 ）表达结果的影响。
被选为跨样本标准化最稳定的参考基因是 、 和 。不同标准化方法对处理对 表达影响的解读存在显著差异。与其他任何标准化方法相比，NORMA-Gene在降低靶基因表达方差方面表现更佳。
我们证明，对于家畜基因表达研究，NORMA-Gene可以提供一种比使用参考基因更可靠的标准化方法，且所需资源更少。
在线版本包含可在10.1186/s12863-025-01345-y获取的补充材料。
