RNAseq of N-starved Vibrio natriegens: parent B
Cell suspensions were incubated in nitrogen-free VMDC175 medium for 2 h at 30C, then chilled on ice and pelleted by centrifugation at 4C. Supernatants were discarded and pellets frozen using dry ice before storing at -80C. RNA was extracted with TRIzolTM (Thermo Fisher) /chloroform. Samples were treated with 4 U Turbo DNase (Invitrogen) in 100 L at 37C for 1 h, then purified using a QIAGEN RNeasy MinElute Cleanup Kit. RNA was quantified using an Agilent 2200 TapeStation at the Indiana University Center for Genomics and Bioinformatics (IU CGB), then submitted to SeqCoast Genomics where rRNA was depleted and libraries prepared using the Illumina Stranded Total RNA Prep Ligation Kit with Ribo-Zero Plus Microbiome and the RNA sequenced on an Illumina NextSeq2000 platform to produce 2x150 bp paired reads.