Based on the three algorithms, we provide a list of genes for cyanobacterial RT-qPCR data normalization. In addition, the minimum number of reference genes required for normalization was determined. The six genes exhibiting less variation in each organism were evaluated in terms of their expression stability using geNorm, NormFinder and BestKeeper. Towards this goal we have selected and analyzed twelve candidate reference genes from three morphologically distinct cyanobacteria grown under routinely used laboratory conditions. However, to obtain reliable RT-qPCR results the use and validation of reference genes is mandatory. For the quantification of gene transcripts real-time quantitative polymerase chain reaction (RT-qPCR) is the standard technique. The use of these organisms as cell factories requires the knowledge of their physiology and metabolism at a systems level. Cyanobacteria are a group of photosynthetic prokaryotes that have a diverse morphology, minimal nutritional requirements and metabolic plasticity that has made them attractive organisms to use in biotechnological applications.
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