Haynes Lab:Notebook/Lab Traning - Pradyumna Kadambi/2013/07/24

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Transformation of DH5α with KAH013 07/24/13

  1. LB plates were taken from fridge and placed in incubator at 37degrees. (By mistake LB-amp plates were selected instead of LB+amp)
  2. KAH013 plasmid was taken from the fridge.
  3. In a 1mL tube, 9μL of dH2O and 1μL of KAH013 were added.
  4. DH5α cells were taken from the freezer and placed in an ice bath.
  5. Waited 10 mins for the DH5α cells to thaw.
  6. In the 1mL tube containing the dH2O and the plasmid, 50μL of DH5α was added.
  7. Waited 10 mins for the bacteria to accept the plasmid.
  8. Pipetted the 60μL contents of the 1mL tube onto the warmed LB-amp plates.
  9. Around 6-10 rolling beads were used to spread the bacteria.
  10. Plates were placed in incubator at 37degrees for 12 hours.

Notes

Because LB-amp plates were used instead of LB+amp plates, a carpet of growth resulted on the plates instead of colonies. So, the process must be done again with LB+amp plates.


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