title: Identification of reference genes suitable for normalization of RT-qPCR expression data in Saccharomyces cerevisiae during alcoholic fermentation.
abstract: Expression data from RT-qPCR (reverse transcription quantitative PCR) needs to be normalized to account for experimental variability among samples caused by differential yields of the transcripts in RNA extraction or in the reverse transcription. The most common method is to normalize against one or more reference genes (RG). We have selected RGs suitable for normalization of RT-qPCR raw data in Saccharomyces cerevisiae during alcoholic fermentation. The RGs were evaluated by three different statistical methods. The suitability of the selected RG sets was compared with ACT1, a commonly used non-validated single RG, by normalizing the expression of two target genes. Expression profiles of the target genes revealed the risk of misleading interpretation of expression data due to an unreliable RG.
authors: CRA-Centro di Ricerca per l'Enologia, Via Pietro Micca 35, Asti, Italy. enricotommaso.vaudano@entecra.it
authors: Enrico
authors: E
authors: Vaudano
authors: Olta
authors: O
authors: Noti
authors: Antonella
authors: A
authors: Costantini
authors: Emilia
authors: E
authors: Garcia-Moruno
mesh_terms: D000426:Alcohol Dehydrogenase / Q000235:genetics; D018076:DNA, Complementary / Q000032:analysis / Q000235:genetics; D005285:Fermentation; D005800:Genes, Fungal; D005954:Glucosephosphate Dehydrogenase / Q000235:genetics; D012331:RNA, Fungal / Q000032:analysis / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D012015:Reference Standards; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics
chemical_list: D018076:DNA, Complementary; D012331:RNA, Fungal; D012333:RNA, Messenger; D029701:Saccharomyces cerevisiae Proteins; C516801:ADH1 protein, S cerevisiae; D000426:Alcohol Dehydrogenase; D005954:Glucosephosphate Dehydrogenase