RNDmutant3h rep3
Overnight cultures of  V. cholerae all six efflux pumps deleted (JB485) were inoculated 1:5000  into fresh AKI medium. Cultures were  grown without shaking for 3hrs at 37?C then cells were collected by centrifugation. Total RNA was isolated using TRIzol (Ambion) according to the manufacturer's intructions The resulting RNA was purified and subjected to DNase treameant using RNeasy Plus Mini Kit (Quiagen), following the manufacturer's protocol. The RNA samples were then quantified using a Nanodrop spectometer (Thermo Fisher) and 1ug of RNA was used to generate RNA libraries using Illumina TruSeq Stranded Total RNA Sample Preparation Kit (Illumina) according to manufacturer's instructions.