Team:Newcastle/IntroductoryLabwork/24 July 2009

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Team Newcastle 2009 iGEM IntroductoryLabSessions.PNG

Introductory Lab Session: 24th July 2009

James and Neil removing most of the 70% ethanol by using pipettes - the DNA pellets will then be dried using the vacuum pump

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

  1. 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 vacuum pump.
  • 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.

June
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[http://2009.igem.org/wiki/index.php?title=Team:Newcastle/IntroductoryLabwork/13_July_2009&action=edit 13] [http://2009.igem.org/Team:Newcastle/IntroductoryLabwork/14_July_2009 14] [http://2009.igem.org/Team:Newcastle/IntroductoryLabwork/15_July_2009 15] [http://2009.igem.org/Team:Newcastle/IntroductoryLabwork/16_July_2009 16] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/IntroductoryLabwork/17_July_2009&action=edit 17] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/IntroductoryLabwork/18_July_2009&action=edit 18] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/IntroductoryLabwork/19_July_2009&action=edit 19]
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[http://2009.igem.org/Team:Newcastle/IntroductoryLabwork/27_July_2009 27] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/IntroductoryLabwork/28_July_2009&action=edit 28] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/IntroductoryLabwork/29_July_2009&action=edit 29] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/IntroductoryLabwork/30_July_2009&action=edit 30] [http://2009.igem.org/wiki/index.php?title=Team:Newcastle/IntroductoryLabwork/31_July_2009&action=edit 31]




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