Selection and validation of reference genes for normalisation of gene expression in ischaemic and toxicological studies in kidney disease.

ABSTRACT: Normalisation to standard reference gene(s) is essential for quantitative real-time polymerase chain reaction (RT-qPCR) to obtain reproducible and comparable results of a gene of interest (GOI) between subjects and under varying experimental conditions. There is limited evidence to support selection of the commonly used reference genes in rat ischaemic and toxicological kidney models. Employing these models, we determined the most stable reference genes by comparing 4 standard methods (NormFinder, qBase+, BestKeeper and comparative ΔCq) and developed a new 3-way linear mixed-effects model for evaluation of reference gene stability. This new technique utilises the intra-class correlation coefficient as the stability measure for multiple continuous and categorical covariates when determining

SUBMITTER: Herath S 

PROVIDER: S-EPMC7241806 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature