840:153g:Projects/project23/2012/10/11

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(Cloning of the OmcF gene)
Current revision (20:13, 16 October 2012) (view source)
(Cloning of the OmcF gene)
 
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==Cloning of the OmcF gene==
==Cloning of the OmcF gene==
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* On Tuesdays, we grew the E.coli that we had received from the Parts Registry; having ordered it the previous Tuesday. This part was vector psB2K3 which contained the promoter BBa_I0500. We plated the bacteria on KanR LB media.
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* On Tuesday, we grew the E.coli that we had received from the Parts Registry; having ordered it the previous Tuesday. This part was vector psB2K3 which contained the promoter BBa_I0500. We plated the bacteria on KanR LB media.
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* On Wednesday we transferred the bacterial cultures into liquid LB media to increase the amount of bacteria to use for extracting the plasmid out again.
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* On Wednesday we transferred the bacterial cultures into liquid LB media to increase the amount of bacteria to use for extracting plasmid DNA.
* On Thursday, we used the GeneJet Plasmid Miniprep Kit to extract our plasmid DNA from the E.coli that we grew on Tuesday and Wednesday
* On Thursday, we used the GeneJet Plasmid Miniprep Kit to extract our plasmid DNA from the E.coli that we grew on Tuesday and Wednesday
* We ran an agrose to gel to verify the existence of plasmid DNA. We used 5 wells in the gel. In wells 1-4 we placed 5 mcL each of our purified plasmid DNA, samples A-D respectively. In well 5 we placed 2 mcL of 100 bp ladder.
* We ran an agrose to gel to verify the existence of plasmid DNA. We used 5 wells in the gel. In wells 1-4 we placed 5 mcL each of our purified plasmid DNA, samples A-D respectively. In well 5 we placed 2 mcL of 100 bp ladder.

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Cloning of the OmcF gene

  • On Tuesday, we grew the E.coli that we had received from the Parts Registry; having ordered it the previous Tuesday. This part was vector psB2K3 which contained the promoter BBa_I0500. We plated the bacteria on KanR LB media.
  • On Wednesday we transferred the bacterial cultures into liquid LB media to increase the amount of bacteria to use for extracting plasmid DNA.
  • On Thursday, we used the GeneJet Plasmid Miniprep Kit to extract our plasmid DNA from the E.coli that we grew on Tuesday and Wednesday
  • We ran an agrose to gel to verify the existence of plasmid DNA. We used 5 wells in the gel. In wells 1-4 we placed 5 mcL each of our purified plasmid DNA, samples A-D respectively. In well 5 we placed 2 mcL of 100 bp ladder.

Our Results: Each prepared plasmid DNA sample showed faint but usable quality DNA.

Image:E coli plasmid DNA.jpg

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