Sequencing of mRNA by NGS - Library preparation

mRNA sequencing (mRNA-Seq), also known as whole transcriptome shotgun sequencing, has become the method of choice for analyzing the transcriptome in different situations such as disease states or biological processes. mRNA sequencing is a very sensitive and accurate way to quantify gene expression.  Furthermore, it allows the identification of known and novel transcriptome isoforms, gene fusions and allele-specific expression. Messenger RNA sequencing provides a complete view of the coding transcriptome that is not limited by the prior knowledge filter.

The most common method is that targeting polyadenylated RNAs as these represent stable mRNAs and therefore not targeted for degradation, which therefore correspond mainly to mRNAs whose purpose is translation into protein. In addition, the selection of polyadenylated RNAs allows the removal of ribosomal RNAs that are not subject to this phenomenon.