TnSeq of P. mirabilis from mouse spleens during bacteremia
Bacterial genomic DNA was isolated from P. mirabilis in the spleens of 10 mice by hexadecyltrimethyl ammonium bromide (CTAB) precipitation. Samples were enriched for transposon insertion junctions as outlined by Goodman et al. (Nat Protoc 2011). TapeStation analysis was used to confirm concentration and purity, and samples were multiplexed and subjected to V4 single end 50 HiSeq-2500 High-Output sequencing. Each lane was spiked with 15% bacteriophage X DNA to overcome low-diversity sequences. Sequencing was performed at the University of Michigan DNA core facility.