TraDIS sequencing of Staphylococcus aureus mutant pool recovered from skin abscess of one mouse (Mouse B)
Transposon mutant library from Staphylococcus aureus USA300 JE2 recovered from a BALB/c mouse skin abscess (Mouse B). Genomic DNA was extracted and prepared following the TraDIS protocol. 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 a Bioanalyzer before sequencing on an Illumina MiSeq using a 50-cycle V2 reagent kit.