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Team:Newcastle/IntroductoryLabwork/24 July 2009
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==Ethanol Precipitation continued== | ==Ethanol Precipitation continued== | ||
| + | [[Image:Team Newcastle 2009 iGEM 24-07-09 IMG 0149.JPG|200px|thumb|Arun and James adding distilled water to the ethanol-free DNA pellets]] | ||
So far we have added sodium acetate to the 2 DNA samples and also added ethanol. The team have also divided the samples into Eppendorf tubes and stored them in the fridge overnight (a replacement for the15 minute incubation-on-ice step). The following was carried out: | So far we have added sodium acetate to the 2 DNA samples and also added ethanol. The team have also divided the samples into Eppendorf tubes and stored them in the fridge overnight (a replacement for the15 minute incubation-on-ice step). The following was carried out: | ||
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* With the supernatant removed the 16 samples were rinsed in 70% ethanol and spun for 15 minutes. The supernatant was discarded (making sure that the DNA pellet was it disturbed) and the tube dried using the Speed Vac machine. | * With the supernatant removed the 16 samples were rinsed in 70% ethanol and spun for 15 minutes. The supernatant was discarded (making sure that the DNA pellet was it disturbed) and the tube dried using the Speed Vac machine. | ||
| - | * The DNA pellets were then resuspended in 50ul of distilled water and stored away in the -80ºC freezer. | + | * The DNA pellets were then resuspended in 50ul of distilled water and stored away in the -80ºC freezer. |
==Conclusion== | ==Conclusion== | ||
Revision as of 17:52, 25 September 2009
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Lab session: 24th July
Introduction
As far as the introductory lab sessions go, we have completed one half of the ethanol precipitation procedure; once completed the procedure will see the plasmid pSB1A2 (containing GFP) and the plasmid pSB1AT3(containing RFP) free from ethanol. This means they can be loaded onto gel and analysed appropriately (i.e. DNA gel electrophoresis)
Practical Outline
- Remove the Eppendorf tubes from fridge and resume the rest of the ethanol precipitation protocol.
Ethanol Precipitation continued
So far we have added sodium acetate to the 2 DNA samples and also added ethanol. The team have also divided the samples into Eppendorf tubes and stored them in the fridge overnight (a replacement for the15 minute incubation-on-ice step). The following was carried out:
- The 16 Eppendorf tubes (8 containing GFP and 8 containing RFP) were spun down for 20 minutes at 13,000 rpm.
- With the supernatant removed the 16 samples were rinsed in 70% ethanol and spun for 15 minutes. The supernatant was discarded (making sure that the DNA pellet was it disturbed) and the tube dried using the Speed Vac machine.
- The DNA pellets were then resuspended in 50ul of distilled water and stored away in the -80ºC freezer.
Conclusion
Both the pSB1A2 plasmid containing GFP and the pSB1AT3 plasmid containing RFP will be analysed by DNA gel electrophoresis on next Monday’s lab session.
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)
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