Team:UCL London/Characterisation
From 2010.igem.org
1. CHARACTERISATION OF NEW PARTS
Our pTAC with RFP reporter was characterised at lab scale by carrying out a successful transformation with a control. By adding transforming the parts into competent cells and adding IPTG to the non-control plate, we expected to see the control's colour remain unchanged whilst that of the other plate to turn red indicating that the IPTG will have induced the activation of the pTAC and thus characterise our part. Obviously, we had hoped to be in a position to characterise the whole genetic circuit but due to time restraints we could complete our circuit.
Check out our parts page to see the parts submitted in more detail [http://partsregistry.org/cgi/partsdb/pgroup.cgi?pgroup=iGEM2010&group=UCL_London&Done=1|UCL_London 2010 iGEM Team Parts].
2. CHARACTERISATION OF EXISTING PARTS/DEVICES AT 1 L FERMENTER SCALE
As part of the Gold criteria which is our target, we had to characterise an existing part, and so we decided to characterise 3 devices from our 2009 parts;
Fermenter 1 - [http://partsregistry.org/wiki/index.php?title=Part:BBa_K239011 UCL_London 2009 iGEM BBa_K239011 PART]
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Fermenter 2 -[http://partsregistry.org/wiki/index.php?title=Part:BBa_K239009 UCL_London 2009 iGEM BBa_K239009 PART]
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Fermenter 3 - [http://partsregistry.org/wiki/index.php?title=Part:BBa_K239015 UCL_London 2009 iGEM BBa_K239015 PART]
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The 3 parts were inoculated into separate 1 L fermenters and we ran the processes in parallel along side a control process. We observed the various stages of the process from the growth to the lag phase, and what we hoped to observe was the progressive change of the colours of the 3 processes into green, as we expected the induction of the GFP protein on addition IPTG causing the induction of the promoter and thus the green proteins release into the culture.
Control Variable
The following video shows the moment of that our iGEM aspirations came true, and we saw green fluorescence;
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K239011 BBa_K239011]
Stress Light GFP Anaerobic Metabolism Detector 2
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K239009 BBa_K239009]
GFP test devise for Spy promoter
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K239015 BBa_K239015] GFP test devise for DegP promoter
From the paste removed from the fermenter vessel following the successful characterisation of the parts, we used it to creatively mark the words iGEM in plates and hence further emphasizing the results that the parts do work.