Selection of Reference Genes for Transcription Studies Considering Co-Regulation and Average Transcriptional Stability: Case Study on Adventitious Root Induction in Olive (Olea europaea L.) Microshoots

Selection of reference genes (RGs) for normalization PCR-gene expression data includes two crucial steps: determination the between-sample transcriptionally more stable genes, and subsequent choosing most suitable as internal controls. Both steps can be carried out through generally accepted strategies, each having different strengths weaknesses. The present study proposes reinforcement gene by integrating analytical revision at critical those procedures. In vitro olive adventitious rooting was used an experimental system. Candidate RGs were ranked according to transcriptional stability several methods. An algorithm one these programs (GeNorm) adapted allow partial automatization RG selection any strategy transcriptional-gene ordering. order choose appropriate set RGs, achieved results analytically revised, with special emphasis on biasing effects such co-regulation. obtained putative sets also tested cases restricted fewer variables. formed H2B, OUB ACT is valid in studies microshoot when comparing treatments, time points assays. Such now available wider target similar biological systems. overall methodology aims constitute a guide general application.