Team:British Columbia/Protocols/ConsortiaFluor

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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.
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.
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Note:  
Note:  

Revision as of 23:46, 3 October 2012

British Columbia - 2012.igem.org
UBC iGEM 2012 protocols


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 a starting 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

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