Transformation of DH5α with KAH013 07/24/13
- LB plates were taken from fridge and placed in incubator at 37degrees. (By mistake LB-amp plates were selected instead of LB+amp)
- KAH013 plasmid was taken from the fridge.
- In a 1mL tube, 9μL of dH2O and 1μL of KAH013 were added.
- DH5α cells were taken from the freezer and placed in an ice bath.
- Waited 10 mins for the DH5α cells to thaw.
- In the 1mL tube containing the dH2O and the plasmid, 50μL of DH5α was added.
- Waited 10 mins for the bacteria to accept the plasmid.
- Pipetted the 60μL contents of the 1mL tube onto the warmed LB-amp plates.
- Around 6-10 rolling beads were used to spread the bacteria.
- 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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