Massively parallel cDNA sequencing (RNA-seq) is a powerful tool for providing an unbiased approach to assess transcript abundance under a variety of conditions. In comparison to microarrays, this technique provides increased resolution and sensitivity and the ability to identify rare transcripts and sRNAs. Here, we describe the sample preparation (based on Illumina technology) used for transcriptomic analysis of V. cholerae cDNA libraries. We describe the entire process from RNA isolation through to the generation of barcoded cDNA libraries ready for sequencing.
