IGEM:UBC/2009/Notebook/Cre excision frequency/2009/08/17

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Assembly ML6 continued=

Assembly ML6 cPCR

  • cPCRed colony A, B and C from spot plate made on Sunday.
    • colony C seemed to have some satellite colonies around it, result must be used with caution and if possible do not use the colony at all.

  • cPCR reveals colony B and C had band at 1300 to 1400bp range. The bands were definitely under 1.5kbp.
    • incorrect band, must redo

Assembly ML6 redone

  • restriction digest started @ 2:36pm to 3:48pm.
  • heat inactivation at 80oC rather than 82oC. Used the old heat block rather than the thermal cycler.
    • heat inactivated from 3:55. was at 80oC for 20minutes.
  • ligation started at 5:45pm.
    • heat inactivated product was left on room temperature for 45 mins and then on ice for another 45 minutes due to time constraints.