Bacterial culture

B. theta strains were stored at –80°C in growth medium mixed in equal volume with 50% glycerol in water. Strains were streaked from glycerol stocks onto BHI/blood agar using plastic inoculating loops. Strains were allowed to grow 24–48 hr in an anaerobic chamber. Single colonies were used to inoculate 4 10 mL aliquots of supplemented BHI broth per strain, and after 24 hr the 10 mL cultures were expanded to 1 L each in glass bottles, producing 4 L total culture volume per strain. Cultures were allowed to grow to stationary phase (24–36 hr) and were pelleted at 3400 × g for 1 hr at 4°C. Pellets were washed in PBS and shipped frozen to the UCSD GlycoAnalytics Core for LPS purification.