Team:Mexico-UNAM-IPN/Horizontal-Transfer/1 October 2008

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TRANSFORMATION OF COMPETENT CELLS JM101

Eniak Hernández

  • Sample (9) BBa_1739201 low copy
  • Sample (11) PUC19 high copy
  1. Thaw the cells on ice for 5 minutes.
  2. Take 50µl of cells and add 5µl of DNA (samples 9 and 11).
  3. Incubate on Ice 20 min.
  4. Heat shock of 40-50 seconds in termoblock at 42°C. DO NOT AGITATE.
  5. Thaw on ice for 2 minutes.
  6. Add 950µl of sterile LB media.
  7. Incubate for 1.5hrs at 37°C at 15rpm.
  8. Plate in petri dishes with LB/amp, 100µl of media. First centrifugate (7min, 3500rpm) and disolve the pellet in 100µl of media, plate. Incubate at 37°C overnight.