Team:Newcastle/Labwork/24 July 2009

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=Lab session: 24th July=
=Lab session: 24th July=
==Introduction==
==Introduction==
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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)
 
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==Practical Outline==
 
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Remove the Eppendorf tubes from fridge and resume the rest of the ethanol precipitation protocol.
 
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==Ethanol Precipitation continued==
 
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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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* The 16 Eppendorf tubes (8 containing GFP and 8 containing RFP) were spun down for 20 minutes at 13,000 rpm.
 
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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.
 
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* The DNA pellets were then resuspended in 50ul of distilled water and stored away in the -80ºC freezer.
 
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==Conclusion==
 
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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.
 
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Revision as of 17:32, 25 September 2009



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