Latest revision as of 03:42, 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.

@@ Line 21: / Line 21: @@
 	<div id="sponsormap"><img align="left" src="https://static.igem.org/mediawiki/2012/d/db/Ubcigemnotebookmenu2.jpg" usemap="#sponsormap" alt="UBC iGEM 2012 protocols"> </div><div id=protocol></html>
 <h1>Consortia Fluorescence Monitoring</h1>
@@ Line 34: / Line 32: @@
 #Incubate in Tecan plate scanner, taking measurements of O.D.600 and fluorescence outputs for time intervals of every 15 minutes for 24 hours.
+<h1>Excitation and Emission Wavelengths of Fluorescent Proteins</h1>
+*EYFP - Excitation: 514nm     Emission: 527nm
+*ECFP - Excitation: 439nm     Emission: 476nm
+*ERFP - Excitation: 584nm     Emission: 607nm
-<h2>Excitation and Emission Wavelengths of Fluorescent Proteins</h2>
-EYFP - Excitation: 514nm     Emission: 527nm
-ECFP - Excitation: 439nm     Emission: 476nm
-ERFP - Excitation: 584nm     Emission: 607nm