GSM2454478: ∆luxO Mucus Replicate 3; Vibrio parahaemolyticus RIMD 2210633; RNA-Seq
Total RNA was extracted from cells obtained by centrifugation at the end of 90 mins using Trizol (Invitrogen, Carlsbad, CA) according to manufacturer's protocol. The RNA samples were then quantified using a Nanodrop spectrophotometer (Thermo Scientific, Waltham, MA). The samples were then treated with Turbo DNase (Invitrogen) according to manufacturer's instructions. For each sample of the wild-type and mutant, RNA samples from two independent cultures were pooled together. Nanodrop quantifications were used to ensure equal representation of RNA from both biological replicates. Then, 3 μg of RNA was used for rRNA depletion using the Ribo-Zero rRNA removal kit for Gram-negative bacteria (Illumina). Libraries for each sample were prepared from 100ng of rRNA-depleted-RNA using the Illumina TruSeq Stranded mRNA kit (Illumina) following standard manufacturer's protocol.