Team:Newcastle/Labwork/10 August 2009

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Today we pepared gfp-rrnb plasmids using midiprep. We followed GenElute's midiprep protocol.
Today we pepared gfp-rrnb plasmids using midiprep. We followed GenElute's midiprep protocol.
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We had two E.coli cultures inoculated with gfp-rrnb left overnight. Although they had teh sampels, we treated them individually.
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We had two E.coli cultures inoculated with gfp-rrnb left overnight. Although the samples were of the same plasmid, we treated them individually as one culture seemed to express the GFP more than the other.
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We used 100ml of each sample aliquoted into two 50ml falcon tubes then carried out the midi prep procedure from the protocol on the 4 tubes, combining our DNA samples at the end.
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In the DNA concentration step we centrifuged the tubes at 16000rpm for 30 minutes before adding ethanol to rinse the DNAs. We then centriguged the tubes again for 10 minutes at 16000rpm.  
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We centrifuged the tubes at 16000rpm for 30 minutes before adding ethanol to rinse the DNAs. We then centriguged the tubes again for 10 minutes at 16000rpm.
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We evaporated the tubes for 5 minutes at the end before resuspending in distilled water.
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We evaporated the tubes fro 5 minutes at the end.
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===Results===
===Results===

Revision as of 22:05, 10 August 2009

Contents

Lab Session 10/08/09

Stochastic Switch Team

Today we pepared gfp-rrnb plasmids using midiprep. We followed GenElute's midiprep protocol.

We had two E.coli cultures inoculated with gfp-rrnb left overnight. Although the samples were of the same plasmid, we treated them individually as one culture seemed to express the GFP more than the other. We used 100ml of each sample aliquoted into two 50ml falcon tubes then carried out the midi prep procedure from the protocol on the 4 tubes, combining our DNA samples at the end. In the DNA concentration step we centrifuged the tubes at 16000rpm for 30 minutes before adding ethanol to rinse the DNAs. We then centriguged the tubes again for 10 minutes at 16000rpm.

We evaporated the tubes for 5 minutes at the end before resuspending in distilled water.

Results

We got good DNA concentrations for both samples. The results are below:

Sample 1

The concentration of DNA: 829.6 ng/uL
The ratio of DNA concentration to protein concentration(260/280): 1.88
The ratio of protein concentration to RNA concentration(260/230): 2.33

Team Newcastle iGEM 2009 08 10 Sample 1.png

Sample 2

The concentration of DNA: 601.1 ng/uL
The ratio of DNA concentration to protein concentration(260/280): 1.90
The ratio of protein concentration to RNA concentration(260/230): 2.11

Team Newcastle iGEM 2009 08 10 Sample 2.png

Chassis team

  • Created germination solution without lysozyme.
  • Borrowed Em from Colin Harwood's lab.
  • Created a new MgCl Stock solution, as old one had bits of matter floating in it.
  • Created KPh stock solution.
  • Autoclaved solutions.
  • Made 250ml LB and 250ml H20, autoclaved.
  • Inoculated LB with Em and Kan, then poured plates.



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