Revision as of 01:22, 4 October 2012

British Columbia - 2012.igem.org

Consortia Fluorescence Monitoring

Grow overnight 5mL cultures of auxotrophs(MetA^-, TrpA^-, and TyrA^-)with their respective fluorescent proteins in LB media and ampicillin (100 mg/mL) .
Spin down cells using a centrifuge (1600 g, 10 min). Pour out LB supernatant.
Wash and resuspend in 5mL M9 media. Spin down cells using a centrifuge (1600 g, 10 min)
Perform step 3 three times.
Resuspend in 5 mL M9 Media.
Measure the respective O.D 600 with a spectrometer.
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).
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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 #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).
 #Incubate in Tecan plate scanner, taking measurements of O.D.600 and fluorescence outputs for time intervals of every 15 minutes for 24 hours.
 <h2>Excitation and Emission Wavelengths of Fluorescent Proteins</h2>
-EYFP - Excitation: 514nm     Emission: 527nm
+*EYFP - Excitation: 514nm     Emission: 527nm
-ECFP - Excitation: 439nm     Emission: 476nm
+*ECFP - Excitation: 439nm     Emission: 476nm
-ERFP - Excitation: 584nm     Emission: 607nm
+*ERFP - Excitation: 584nm     Emission: 607nm