Sequencing for Tn10 deletion
Genomic DNA was extracted from overnight cultures of the SSuT6 and the SSuT6 tetracycline-susceptible strain using an UltraClean Microbial DNA isolation kit (Mo Bio Laboratories,Inc. Carlsbad, CA). DNA was sheared using a M220 Focused-ultrasonicator (Covaris, Woburn, Massachusetts) before library generation using the NugenOvation Ultralow System V2 1-16 (XXX, San Carlos, CA). An Illumina MiSeq instrument was used to generate sequences by using a . Raw data was trimmed using Trim Galore! and assembled using SPAdes genome assembler. The Sequence Search Tool for Antimicrobial Resistance (SSTAR) was used to identify where tet(B) and associated Tn10 genes was located in the SSuT6 sequences. The absence of this transposon was noted in the tetracycline-sensitive strain, corresponding to complete excision of the Tn10 region. No other mutations were evident from alignments (true?) based on (what software or analysis?). The sequenced tetracycline-sensitive strain was designated Tn10.