IGEM:IMPERIAL/2006/LabCalendar/2006-7-28

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Mini-prep attempt number 2

  • Parts 2H (I13033), 16P (J04500), 5M (C0060)cultured previous day mini-preped.
  • DNA extracted and digested.
  • Gel elecrophoresis resulted in the following bands.




The four lanes show DNA markers, 2H, 5M and 16P from left to right. Individual band sizes correspond to their respective plasmid and parts sizes as listed in registry.
Parts successfully cultured in bacteria. Ready for ligations.

Ligations

  • Re washed DNA with gene clean wash protocol
  • Electroporated the following into competent cells
    • I13504(insert) into C0261(vector)
    • I13504(insert) into I13033(vector)
    • C0062(insert) into B0034(vector)
    • 1I from parts registry

Note that the gene clean wash to purify the DNA from ligase needs to be the altered version of protocol

  • Add equal volume of 6M NaI instead of 500 uL
  • Skip 5 minute incubation in step 2
  • Elute in 20 uL in step 10
  • Remove 15 uL of supernatant in step 12
  • Respin and take out 10 uL from that to a new eppendorf tube...this will be the DNA we use for the electroporation

To do over weekend

  • Dr. Mann will collect plates from the 37C incubator on Saturday
  • Miniprep parts electroporated today on Sunday
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