Team:Newcastle/Labwork/22 September 2009

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Lab Work - 22/09/09

Stochastic switch team

Today we transformed E.coli JM109 cels with the overnight ligation product from yesterday using the promega protocol. We used the following controls:

  • Tube 1: Backbone + insert + ligase (proper ligation)
  • Tube 2: Backbone + ligase (control for background ligation level)
  • Tube 3: Backbone + insert (control for ligase effectiveness)

These were left in the incubator for an hour and then plated out (2 plates per tube 200ul and 50ul) onto Amp + Tet LB plated and put in the 37C incubator overnight.

Today again we redid the digests of the sac miniprep however again there was no DNA present.




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