biostudies-literatureUnknown6Martins PKReal-time PCR (RT-qPCR) expression analysis is a powerful analytical technique, but reliable results depend on the use of stable reference genes for proper normalization. This study proposed to test the expression stability of 13 candidate reference genes in Setaria viridis, a monocot species recently proposed as a new C4 model plant. Gene expression stability of these genes was assayed across different tissues and developmental stages of Setaria and under drought or aluminum stress. In general, our results showed Protein Kinase, RNA Binding Protein and SDH as the most stable genes. Moreover, pairwise analysis showed that two reference genes were sufficient to normalize the gene expression data under each condition. By contrast, GAPDH and ACT were the least stably expressed genes tested. VScientific reports28348https://www.ebi.ac.uk/biostudies/studies/S-EPMC4913262biostudies-literatureSelection of reliable reference genes for RT-qPCR analysis during developmental stages and abiotic stress in Setaria viridis.PMC4913262Kobayashi AKRibeiro APde Souza WRBasso MFda Cunha BAMolinari HBVinecky FMartins PKMafra VfalseSelection of reliable reference genes for RT-qPCR analysis during developmental stages and abiotic stress in Setaria viridis.Real-time PCR (RT-qPCR) expression analysis is a powerful analytical technique, but reliable results depend on the use of stable reference genes for proper normalization. This study proposed to test the expression stability of 13 candidate reference genes in Setaria viridis, a monocot species recently proposed as a new C4 model plant. Gene expression stability of these genes was assayed across different tissues and developmental stages of Setaria and under drought or aluminum stress. In general, our results showed Protein Kinase, RNA Binding Protein and SDH as the most stable genes. Moreover, pairwise analysis showed that two reference genes were sufficient to normalize the gene expression data under each condition. By contrast, GAPDH and ACT were the least stably expressed genes tested. V2016-01-01T00:00:00Z2016 Jun2026-05-05T07:22:43.414Z2019-03-27T02:16:26ZS-EPMC49132622732167510.1038/srep28348