Collection Summary:

The wildtype S. aureus, nos mutant, SrrAB mutant, nos SrrAB double mutant, and complement strains will be grown in 40 ml trytic soy broth lacking dextrose for 4 hours. At 4 hours growth, all 40 ml was isolated, centrifuged at 4500 rpms and 4C for 10 minutes and then placed on ice. 1 ml of supernatant was removed in duplicate and saved as "supernatant". The rest of the supernatant was discarded and pellets were resuspended in 2 ml PBS, centrifuged at 4500 rpms for 3 minutes (4C). Supernatant was discarded and samples were again resuspended in 2 ml PBS. These samples were split into 2 separate 1 ml aliquots (for metabolite and protein determination) and frozen with liquid nitrogen. Separate media only samples of Tryptic Soy Broth without dextrose were isolated for each day (3 total) Samples were stored at -80C. A total of 39 samples (18 whole cell, 18 supernatant, and 3 media) were isolated.