IGEM:IMPERIAL/2006/LabCalendar/2006-8-12

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Contents

Minipreps to check ligations of J37015 and J37016

The following parts were mini-preped:

  • L1, L2, L3, L4
  • M1, M2, M3, M4
  • R1, R2, R3, R4
  • V1, V2, V3, V4
  • Z1, Z3, Z4, Z5

Below is the one of the gels from the miniprep: The bands run (from left to right)

  • M1, M2, DNA ladder, R4, R3, R2, R1

Below is the second minprep gel containing (from left to right) :

  • Z5, Z4, Z3, Z1, DNA ladder, V4, V3, V2, V1, L4, L3, L2, L1



Problems

  • Gels overran so no conclusive evidence could be obtained
  • There were unexpected bands
    • This could be due to the fact that the cultures being minipreped were old
  • Could not run M3, M4 since we had no more enzyme for the digest left

Actions

  • Re-cultured the colonies which appear to have worked for re-minipreping these were
    • J37015
      • L 2,3,4
      • M 1,2
    • J37016
      • V 2,3,4
      • R 3,4

These have to be minipreped tomorrow and then digests have to be done on Monday

Maxiprep of 2H

successful

Ligation

  • Ligation of 12D->2H and electroporated
  • Cultured on LB Amp Plates and grown overnight

Gene Clean & Electroporation

  • Cre 1, 2, 3, 4
  • AiiA
  • AiiA, Cre 1, 2, and 4 were electroporated, cultured on LB Amp plates and grown overnight
    • Cre 3 sparked on electroporation, did not re-attempt since we already have three
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