title: Illumina MiSeq paired end sequencing; Identification of the teixobactin-induced CroRS regulon in Enterococcus faecalis using RNA-seq.
design: Identification of the teixobactin-induced CroRS regulon in Enterococcus faecalis using RNA-seq.
library_construction_protocol: 30 ml cultures were mixed with equivolume 3:2 glycerol saline and harvested by centrifugation at 10,000 x g for 15 minutes at -10 degrees celcius. Cells were resuspended in 1ml 1:1 glycerol saline and stored at -80 degrees celcius. Enterococcus faecalis wild-type and the croRS deletion mutant were grown in biological triplicate in BHI broth to an OD600 of 0.5 at 37 degrees celcius and 130 rpm. Following growth to an OD600 of 0.5, cultures were split in two, with half treated with teixobactin (0.5 ug/ml) and the other half untreated and grown for a further 1 h. TRIzol/chloroform RNA extraction, followed by RNA clean and concentrate using a Zymo RNA cleand and concentrate kit. RNA libraries were prepped using the Zymo-Seq Ribofree Total RNA-Seq Library kit.