"
Team:DTU-Denmark/Achievements/Interlab study
From 2014.igem.org
- Home
- Overview
- Methods
- Achievements
- Team
At the Giant Jamboree, we received a gold medal and an Interlab Study Award. We also won the Overgrad Best Parts Collection.
Construct strains
We created multiple strains expressing GFP from different promoters from the Andersen Library. 12 out of the 15 promoters we intended to use were successfully transformed into DH5α These 12 constructed strains were applied in the further analysis of the relatively promoter strength. J23111, J23117 J23109 was not succesfully cloned into E. coli. We did not detect any fluorescence signal from these strains and it was confirmed from the sequencings. we excluded these strains. E0240: Uden promoter GFP E0206: Canamycin I20260.Fluorescence measurement on cultures
Fluorescence was measured on the O/N cultures with our constructed strains in the BioLector. Biomass and fluorescence were measured during growth. Fluorescence signal through the growth was normalized by dividing by OD600. The average of the fluorescence is illustrated in the bar chard below. Together with the expected values relative to the J23100 promoter. Orange is the expected values and gray indicates our measured fluorescence signal.
Single cell measurements
2 mL LB (0.5% NaCl) was inoculated with cells from a plate and grown overnight. The cultures were washed and resuspended in PBS. The cell suspensions were diluted in PBS to a concentration where the flow cytometer could detect 1000-1500 events per second.For more detailed information on the instruments used, settings and the measured quantities see the filled out interlab form.
Technical University of Denmark
Department of Systems Biology
Søltofts Plads 221
2800 Kgs. Lyngby
Denmark
+45 45 25 25 25
dtuigem14@gmail.com .
+45 45 25 25 25
dtuigem14@gmail.com . 
