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RNA-seq

Transcriptome Analysis is the study of the transcriptome, of the complete set of RNA transcripts that are produced by the genome, under specific circumstances or in a specific cell, using high-throughput methods.

Brief outline of the workflow

1. bulk RNA is extracted from the sample and the desired RNA is selected (sample preparation)

2. the selected RNA is copied into stable double-stranded copy DNA (library construction)

3. the ds cDNA is then sequenced using various sequencing methods

4. the sequences obtained can are aligned to reference genome sequences, available in data banks, to identify which genes are transcribed. This type of analysis provides a quantification of the expression levels for the transcribed genes. Alternatively, RNA-seq can be used to identify alternative splicing, novel transcripts, and fusion genes, following a new transcript discovery approach.

The complete workflow of RNA-seq consists of: (1) experimental design; (2) sample and library preparation; (3) sequencing; and (4) data analysis. You will find a general explanation of each step in the following video.

For a deeper understanding of the RNA-seq technology and its applications follow these links:

https://www.intechopen.com/books/applications-of-rna-seq-and-omics-strategies-from-microorganisms-to-human-health/rna-seq-applications-and-best-practices