Team:Newcastle/IntroductoryLabwork/3 July 2009
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- | [[Image:Team Newcastle iGem 2009 - Lab 03-07-09 no 2.JPG|thumb|right|Jane and Jess preparing serial dilutions]] | + | ==Introductory Lab session: 3rd July 2009== |
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+ | [[Image:Team Newcastle iGem 2009 - Lab 03-07-09 no 2.JPG|200px|thumb|right|Jane and Jess preparing serial dilutions]] | ||
Today we were transforming DNA using Phil's protocol, however we used the following changes: | Today we were transforming DNA using Phil's protocol, however we used the following changes: | ||
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====Our serial dilutions were prepared as follows:==== | ====Our serial dilutions were prepared as follows:==== | ||
- | [[Image:Team Newcastle iGem 2009 - Lab 03-07-09 no 5.JPG|thumb|right|Spreading the diluted cells across the agar surface using sterile glass beads]] | + | [[Image:Team Newcastle iGem 2009 - Lab 03-07-09 no 5.JPG|200px|thumb|right|Spreading the diluted cells across the agar surface using sterile glass beads]] |
* Using the waiting times within our procedure we set up 6 labelled eppendorf tubes with 900µl of LB, ready for the series dilution. | * Using the waiting times within our procedure we set up 6 labelled eppendorf tubes with 900µl of LB, ready for the series dilution. | ||
* We took 100µl of our incubated cells and put them in tube 1, mixed the solution briefly and then proceded to take 100µl of the diluted cells and tranfer them along the line of eppendorfs, resulting in eppendorfs containing 10-1 to 10-5 solutions for both the control and 'transformed' cells. | * We took 100µl of our incubated cells and put them in tube 1, mixed the solution briefly and then proceded to take 100µl of the diluted cells and tranfer them along the line of eppendorfs, resulting in eppendorfs containing 10-1 to 10-5 solutions for both the control and 'transformed' cells. | ||
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Latest revision as of 22:22, 12 October 2009
Introductory Lab session: 3rd July 2009
Today we were transforming DNA using Phil's protocol, however we used the following changes:
- We added 3µl of DNA to our cells.
- We plated out 100µl of our dilutions.
- We incubated our transformed cells for the full 60 mins.
- We used the ampicillin resistance plasmid '435'
Procedure
- We followed the protocol using sterile technique where possible, as well as wearing gloves to prevent contamination of the DNA.
- Glass tops such as LB jars can be run under the flame, however plastic eppendorfs cannot so working around the flame is sufficient.
- We found it easier if the person pipetting held the eppendorf as they can see what they're doing better.
- Prof. Aldridges lab uses sterile beads to plate ourt rather than plastic spreaders, these are a bit fiddly so it may take a while to get used to tipping the right amount out!!
Our serial dilutions were prepared as follows:
- Using the waiting times within our procedure we set up 6 labelled eppendorf tubes with 900µl of LB, ready for the series dilution.
- We took 100µl of our incubated cells and put them in tube 1, mixed the solution briefly and then proceded to take 100µl of the diluted cells and tranfer them along the line of eppendorfs, resulting in eppendorfs containing 10-1 to 10-5 solutions for both the control and 'transformed' cells.
- We plated all 6 transformed E.coli solutions onto LB+amp agar plates, and the controls we plated onto both LB+amp plates(to check for contamination) as well as plain LB plates (in order to later calculate transformation frequency).
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News
Events
- 20 – 21 June 2009 - Europe workshop (London)
- 23 – 24 June 2009 - UK iGEM meetup (Edinburgh)
- 23 October Practice Presentation (Newcastle)
- 23 October T-shirts are ready
- 27 October Practice Presentation (Sunderland)
- 27 October Poster is ready
- 30 October – 2 November 2009 - Jamboree (Boston)
Social Net
- Newcastle iGEM Twitter
- [http://www.facebook.com/home.php#/group.php?gid=131709337641 Newcastle on Facebook]
- [http://www.youtube.com/user/newcastle2009igem Newcastle Youtube Channel]