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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 (100mg/mL) .

2. Spin down cells using a centrifuge (1600g, 10min). Pour out LB supernatant.

3. Wash and resuspend in 5mL M9 media. Spin down cells using a centrifuge (1600g, 10min)

4. Perform step 3 three times.

5. Resuspend in 5mL M9 Media.

6. Measure the respective O.D 600 with a spectrometer.

7. Inoculate appropriate co-culture of auxotrophs (each with an O.D.600 of 0.05) in a 96 well plate containing 200uL M9 with ampicillin (100mg/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.

Note:

EYFP - Excitation: 514nm Emission: 527nm ECFP - Excitation: 439nm Emission: 476nm ERFP - Excitation: 584nm Emission: 607nm