TRANSCRIPTOME ANALYSIS USING RNA-SEQ FROM EXPERIMENTS WITH AND WITHOUT BIOLOGICAL REPLICATES: A REVIEW

The discovery of nucleic acids opened new frontiers of knowledge, enabling researchers to access an enormous amount of data, through large-scale sequencing methodologies and bioinformatics tools. Amongst these new possibilities, RNA-Seq has been used to identify and quantify RNA molecules. To obtain more accurate biological responses from RNA-Seq data some questions should be considered such as experimental design, type of synthesized library, size of the fragments generated, number of biological replicates, depth, and coverage of the sequencing, species genome availability, and, the choice of software to properly perform the computational analyzes. Accurate bioinformatics analyzes allow the selection of genes with a lower error rate, increasing the validation assertiveness via RT-qPCR and thus, reducing costs. The objective of this review was to present the analysis stages of RNA-Seq data, from experimental design to system biology, considering relevant points, as well as to pointed out some software currently available to carry these analyzes out. Besides, with this review, we aimed to help the academic community to understand all steps and biases involved in RNA-Seq data analysis, from experiments with or without biological replicates.