RNA Preparation and RNA-Seq Bioinformatics for Comparative Transcriptomics

Overview

Journal Methods Mol Biol

Specialty Molecular Biology

Date 2023 Aug 29

PMID 37642840

Authors

Antonio Rodriguez-Garcia

Alberto Sola-Landa

Carlos Barreiro

Affiliations

Soon will be listed here.

Abstract

The principal transcriptome analysis is the determination of differentially expressed genes across experimental conditions. For this, the next-generation sequencing of RNA (RNA-seq) has several advantages over other techniques, such as the capability of detecting all the transcripts in one assay over RT-qPCR, such as its higher accuracy and broader dynamic range over microarrays or the ability to detect novel transcripts, including non-coding RNA molecules, at nucleotide-level resolution over both techniques. Despite these advantages, many microbiology laboratories have not yet applied RNA-seq analyses to their investigations. The high cost of the equipment for next-generation sequencing is no longer an issue since this intermediate part of the analysis can be provided by commercial or central services. Here, we detail a protocol for the first part of the analysis, the RNA extraction and an introductory protocol to the bioinformatics analysis of the sequencing data to generate the differential expression results.

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