Team:Evry/Interlab Study/08-21-2014

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Contructions PSB1C3 with I20260, J23101-E1010 and K823012-E1010


Transformations of registry parts (BBa_K823012 and J23101) were performed to obtain colonies with plasmid containing parts for future amplifications.
The transformation was performed on DH5 alpha E. coli, as followed:

  1. Remove E. coli competent tubes from -80°C and keep it on ice
  2. Add 3 µl of template (here solubilized plasmids from the registry distribution kit) and mix gently
  3. Incubate 10 minutes on ice
  4. Perform an heat shock 30 seconds at 42°C
  5. Incubate 2 minutes on ice
  6. Add 2 ml of LB medium and incubate 60 minutes at 37°C with an agitation at 200 rpm
  7. Plate 200 µl of BBa_K823012 or BBa_J23101 on a chloramphenicol LB agar plate.
  8. Incubate plate overnight at 37°C

Aug 21