Team:British Columbia/Protocols/ConsortiaFluor

From 2012.igem.org

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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).
#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.
#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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<h2>Excitation and Emission Wavelengths of Fluorescent Proteins</h2>
<h2>Excitation and Emission Wavelengths of Fluorescent Proteins</h2>
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EYFP - Excitation: 514nm    Emission: 527nm  
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*EYFP - Excitation: 514nm    Emission: 527nm  
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ECFP - Excitation: 439nm    Emission: 476nm
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*ECFP - Excitation: 439nm    Emission: 476nm
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ERFP - Excitation: 584nm    Emission: 607nm
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*ERFP - Excitation: 584nm    Emission: 607nm

Revision as of 01:22, 4 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 (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