Team:UCL London/Characterisation

From 2010.igem.org

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[http://partsregistry.org/wiki/index.php?title=Part:BBa_K239015'''UCL_London 2009 iGEM BBa_K239015 PART''']===
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===Fermenter 3 - [http://partsregistry.org/wiki/index.php?title=Part:BBa_K239015'''UCL_London 2009 iGEM BBa_K239015 PART''']===
  GFP test devise for Deg P promoter
  GFP test devise for Deg P promoter
[[Image:UCL-CHARAC15.png|800px|left]]
[[Image:UCL-CHARAC15.png|800px|left]]

Revision as of 00:57, 28 October 2010

UCL IGEM 2010

RETURN TO IGEM 2010

1. CHARACTERISATION OF NEW PARTS

Our pTAC with RFPBBa_K301001 reporter was characterised at lab scale by carrying out a successful transformation with a control.

Ucl-Part-BBa-K301001.png


 


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).


The following is the manual for the construction of BBa_K301000

Check out our parts page to see the parts submitted in more detail 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 - UCL_London 2009 iGEM BBa_K239011 PART

Stress Light GFP Anaerobic Metabolism Detector: mNARK promoter 
UCL-CHARAC11-1.png
UCL-CHARAC11-2.png




...


 

Fermenter 2 -UCL_London 2009 iGEM BBa_K239009 PART

GFP test devise for Spy promoter

UCL-CHARAC9.png ...



 

Fermenter 3 - UCL_London 2009 iGEM BBa_K239015 PART

GFP test devise for Deg P promoter
UCL-CHARAC15.png




 

Images from fermentation chamber

MNARK Infors 05.jpg
MNARK Infors 02.jpg


 

The image on the left is that of the control, while that on the 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 UCL iGEM in plates and hence further emphasizing the results that the parts do work.

UCL-MNARK paste.jpg
 


MNARK IGEM plus control 02.jpg
MNARK IGEM plus control 03.jpg
 

These images further conclude that we succesfully characterised our mNark Promoter as both images have both a control with no observed dye and one showing the green dye, with which we crafted the words UCL and iGEM as shown above.


3. Charactisation of Cambridge Part:BBa_K274003

K274003 Diagnostic Gel .jpg

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 Cambridge Part Review

 



 

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