1 mL of a ML5 transposon library aliquot grown in LB+ 50µg/mL kanamycin to an OD600 of ~1.0  and saved at -80 ˚C was inoculated into 25 mL M9 + 20 mM D-glucose (125 mL baffled flask) and grown to an OD600 of ~1.0 (shaking 225 RPM) prior to pelleting and storing at -80 ˚C