title: Tn-seq of Staphylococcus aureus mutant library input control 3
design: Mice were intranasally infected with 20,000,000 cfu of the library and 24 h later lungs were removed and homogenized through 40 m filters. Lung homogenates were then outgrown for 5 h in LB at 37C. The library was prepared for sequencing following the protocol of [Grosser MR, Paluscio E, Thurlow LR, Dillon MM, Cooper VS, Kawula TH, et al. Genetic requirements for Staphylococcus aureus nitric oxide resistance and virulence. PLoS Pathog. 2018;14(3):e1006907] using a Covaris sonicator that generated 300-700 bp sized DNA fragments.