IGEM:UBC/2009/Notebook/Cre excision frequency/2009/08/15

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Assembly ML6

ML6 digestion

  • Restriction digest started @ 10:14am to 11:15am.
  • heat inactivated @ 11:16 to 11:38.

ML6 ligation

  • ligation (16oC)@ 11:40 to 2:40pm.
    • was left in thermal cycler at 4oC until 3:50pm.

ML6 transformation

  • ligated product introduced into DH5a competent glycerol cell @ 4:08pm
  • on ice from 4:08 to 4:38pm
  • heat shocked for 1 min and on ice for 2 mins(in span of 4:39 to 4:42)
  • recovery from 4:42 to 6:44pm
  • spread plated @ 6:47pm
    • plate was left in Biosafety cabinet to dry for 20 minutes.
    • incubation started at 7:11pm.

Other stuffs

  • made LB agar and autoclaved it.


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