GSM7900515: RNAseq_VP882 wt 60 min replicate i; Vibrio cholerae; RNA-Seq
RNA was isolated using Tri-reagent. Prepared RNA was digested for DNA contaminations (TurboDNase treatment) and cleaned up using standard nucleic acid precipitation procedures. Depletion of ribosomal RNA was performed by rRNA-capturing biotinylated oligos. Binding to magnetic streptavidin-beads removed rRNA. Depletion was verified by Bioanalyzer.  doi.org/10.1128/mBio.00010-20 The rRNA depleted RNAs samples were fragmented using magnesium fragmentation module (NEB). cDNA was synthesized by using the NEBNext® Multiplex Small RNA Library Prep Set for Illumina® (NEB) as specified by the manufacturer. cDNA libraries were column-based purified and cleaned left-sided using the Agencourt AMPure XP kit (Beckman Coulter Genomics). The quality and quantity of cDNA libraries was measured by Bioanalyzer and Qubit, respectively.