title: RNA-Seq of B.subtilis_py79: wt infected with daimXdyosL phi3T phage, rep1
design: Total RNA was extracted from cells using a High Pure RNA Isolation kit (Roche). To this end, strains were grown overnight in LB at 37 with shaking at 220 RPM then diluted by a factor of 1:100 into fresh LB media. Upon reaching OD600 = 0.3 cells of wild-type, mazEF and mazEF;Pxyl-mazF (induced after dilution with either 0.001%, MazF+, or 0.01%, MazF++, w/v xylose) were infected with 3T; aimX; yosL. MazF++ and mazEF were also infected by 3T(vir); aimX; yosL. All infections were done at a MOI of 10 and then incubated with shaking for 20 minutes (37, 220 RPM). Samples were centrifuged for 5 minutes at 2000 RCF and pellets were flash-frozen in liquid nitrogen. Total RNA was extracted from cells using a High Pure RNA Isolation kit (Roche). Before proceeding to library preparation, RNA quantity and quality were validated via Nanodrop, Qubit, and 4200 TapeStation. rRNA removal was conducted using the NEBNext rRNA Depletion Kit (Bacteria) (NEB, #E7860L). Paired-end RNA-seq libraries were generated using the NEBNext Ultra Directional RNA kit (NEB, #E7420S).  Paired-end sequencing was performed on an Illumina NextSeq500 at the ZABAM Instrumentation and service center at Tel Aviv University.