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Revision as of 17:29, 27 October 2010

UCL IGEM 2010

1. CHARACTERISATION OF NEW PARTS

Our pTAC with RFP reporter was characterised at lab scale by carrying out a successful transformation with a control.

IPTG induction of the reporter device BBa_K301001 was demonstrated in a lacI host strain with 0.1mM IPTG (panel B). A lac promoter device was tested as positive control (panel C).

As predicted, IPTG induction was not observed when our new device resided in a lacI negative host strain (panel A) or when the pTac biobrick was omitted (panel D).

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_K239015UCL_London 2009 iGEM BBa_K239011 PART]

Stress Light GFP Anaerobic Metabolism Detector: mNARK promoter

...

Fermenter 2 -[http://partsregistry.org/wiki/index.php?title=Part:BBa_K239009 UCL_London 2009 iGEM BBa_K239009 PART]

GFP test devise for Spy promoter

...

Fermenter 3 - [http://partsregistry.org/wiki/index.php?title=Part:BBa_K239011 UCL_London 2009 iGEM BBa_K239015 PART]

GFP test devise for Deg P promoter

Images from fermentation chamber

The image on the left is that of the control, while that on tge right showing the characeterised mNark promoter fluorescent green.

The following video shows the moment of that our iGEM aspirations came true, and we saw green fluorescence;

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.

3. Charactisation of Cambridge Part:BBa_K274003

E. coli turned dark green after transformation of this plasmid. This is unexpected as there is no promoter upstream of the operon.

Diagnostic DNA gel was run to test all 4 restriction sites. BBa_K274003 was cut with XbaI, PstI, EcoRI & SpeI, XbaI & PstI. A gel picture is shown as below.

This plasmid did not cut with Xba I restriction site in our hands. As such we could not use it to assemble after other parts.

Click on the following link for the [http://partsregistry.org/Part:BBa_K274003:Experience#User_Reviews Cambridge Part Review]

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 {{:Team:UCL_London/clear}}
+'''''Images from fermentation chamber'''''
-'''''Control Variable'''''                                  '''''Test Variable'''''
 [[Image:MNARK Infors 05.jpg|300px|left]]   [[Image:MNARK Infors 02.jpg|300px|left]]
 {{:Team:UCL_London/clear}}
+The image on the left is that of the control, while that on tge right showing the characeterised mNark promoter fluorescent green.
 The following video shows the moment of that our iGEM aspirations came true, and we saw green fluorescence;
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Team:UCL London/Characterisation