LABORATORY OF INTESTINAL MICROBIOMICS
E. coli never takes a vacation
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Stocks for plates
Antibiotic concentrations
1000X Streptomycin 100 mg/mL
200X Nalidixic Acid 10 mg/mL
1000X Chloramphenicol 30 mg/mL (EtOH)
1000X Kanamycin 40 mg/mL
1000X Ampicillin 100 mg/mL
200X Rifampicin 10 mg/mL (DMSO)
200X Tetracycline 10 mg/mL (EtOH)
Everyone has a job
Lab Duties
1. A la carte
2. TAE, MilliQ, maintain filters
3. Biohazard
4. Sterilize tubes, flasks, cages, etc.
5. Stock gloves, empty trash
6. Common media
7. Declutter
8. Clean glassware, etc.
9. Prepare stuff for autoclaving
10. Organize chemicals, check drawers, inventory
Notes to a competitive colonization
Standard Mouse Colonization
Allow mice to acclimate for one week. Begin streptomycin-water (5 g/L) at least 2 days before the experiment.
1. 2 days prior to colonization, streak strains onto LB
2. 1 day prior, inoculate strains into broth cultures.
3. 14-18 hours before colonization, remove streptomycin-water and food for overnight.
4. Prepare 10^5 CFU of each strain combined in 20% sucrose in a sterile bottle cap for mice.
5. Perform viable counts on the inoculum. Return food and streptomycin-water to mice after consuming inoculum.
6. Collect first sample at 5 hr and serially dilute in 1% tryptone (1 g feces/10 mL tryptone).
7. Repeat step 6 at 24 h, 72 h, and every other day for 15 days.
8. Count plates 24-48 h after plating and record CFU.