TraDIS of Staphylococcus aureus transposon mutant library grown in blood (R3)
Fragmented DNA was processed to blunt-ended and dA-tailed using NEBNext modules. Transposon-containing fragments were enriched using the splinkerette method with SpIA5 adaptors and a quick ligation module. Post-ligation, transposon sequence fragments were enriched via PCR with specific primers. DNA fragments were cleaned and size-selected using AMPure XP beads after each library preparation step. Libraries were quantified and verified using SYBR Green qPCR and Bioanalyzer before sequencing with a MiSeq 50-cycle V2 reagent kit following the TraDIS protocol