GSM4476515: rneTS 30 rep1; Vibrio cholerae; RNA-Seq
Total RNA was isolated using the Hot Phenol method as described previously (Fröhlich et al., 2018). Prepared RNA was DNAse digested and cleaned up using standard nucleic acid precipitation procedures. cDNA libraries were prepared by vertis Biotechnology AG: total RNA samples were poly(A)-tailed and 5'PPP structures were removed using RNA 5'Polyphosphatase (Epicentre). An RNA adapter was ligated to the 5' monophosphate and first-strand cDNA synthesis was performed using an oligo(dT)-adapter and M-MLV reverse transcriptase. The resulting cDNAs were PCR-amplified, purified using the Agencourt AMPure XP kit (Beckman Coulter Genomics).