Tn-Seq Survival Profiling

A Tn5 transposon insertion library generated in a thyA- strain was selected in thymidine-free media and insertions were mapped and sequenced at different time points to look for changes in survival caused by gene disruptions Overall design: The library was thawed and pre-grown for 2h in high thymidine, washed twice in 1x phosphate buffered saline (PBS), and placed in thymidine-free media. Samples were collected at three time points during the selection: “Start”, “0h”, and “3h”.