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Consortia Fluorescence Monitoring

1. Grow overnight 5mL cultures of auxotrophs(MetA^-, TrpA^-, and TyrA^-)with their respective fluorescent proteins in LB media and ampicillin (100 mg/mL) .
2. Spin down cells using a centrifuge (1600 g, 10 min). Pour out LB supernatant.
3. Wash and resuspend in 5mL M9 media. Spin down cells using a centrifuge (1600 g, 10 min)
4. Perform step 3 three times.
5. Resuspend in 5 mL M9 Media.
6. Measure the respective O.D 600 with a spectrometer.
7. Inoculate appropriate co-culture of auxotrophs (each with a starting O.D.600 of 0.05) in a 96 well plate containing 200 μL M9 with ampicillin (100 mg/mL).
8. Incubate in Tecan plate scanner, taking measurements of O.D.600 and fluorescence outputs for time intervals of every 15 minutes for 24 hours.

Excitation and Emission Wavelengths of Fluorescent Proteins

EYFP - Excitation: 514nm Emission: 527nm

ECFP - Excitation: 439nm Emission: 476nm

ERFP - Excitation: 584nm Emission: 607nm