RNA-Seq for VPA0519 mutant
A total of 10 microliter of glycerol stock was inoculated into 5 milliliter of 2.5 percent heart infusion broth and incubated at 37 degree with shaking for 13 hours until reaching the stationary phase. The culture was diluted 40-fold in 2 milliliter of HI broth, mixed thoroughly, and spread evenly onto a 6-centimeter diameter HI agar plate using 200 microliter of the dilution. Plates were incubated statically at 37 degree for 6 hours. Bacterial cells were collected in triplicate from HI agar plates for total RNA extraction and RNA-seq.