IGEM:IMPERIAL/2006/LabCalendar/2006-8-16

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Contents

12D->2H

  • The gel that was run yesterday was inconclusive as to whether the DNA has the problem or the enzyme.
  • We will run 7A using the same procedure to see if it is cutting the plamid at all. We are again using the same enzymes with yellow buffer
  • SpeI & PstI were able to successfully cut 7A, so it is not a problem with the enzyme!
  • We will try re-electroporating part 2H again using the DNA from the parts registry to see if we can get it to work tomorrow


Reculture J37016

    • Plates were quite good and we managed to isolate single colonies
  • We took a total of 4 cultures to miniprep tomorrow or later this afternoon to see if we get what we expect.

Ligation of 6B+RS+12D+24A

  • The insert and vector were removed from the overnight gel
  • 20 uL ligation performed

Electroporations

  • J37015RS (6B->RS->12D->24A)
  • 1M B0021 removed from iGEM plate
  • 4G S01656 removed from iGEM plate 2, grown on kanamycin plate
  • 2H I13033 re-electroporated to see if the SpeI site has mutated on the gene
  • Didn't bother with adding LB, since we only have LB Amp and no Kanamycin. So just plated the electrocompetent cells straight onto plate. Hopefully this doesn't affect growth.

Set up Maxipreps

  • 6B F2620 (ran out of maxiprep)
  • 1I B0015 (remaxiprep, since whereabouts of previous maxiprep unknown)

To do tomorrow

  • Maxiprep parts above
  • Reculture the electoporations into liquid media
  • Ligate 1I and 9G to make part 2H, in case 2H fails yet again
  • Miniprep the parts from electroporation, if possible
  • Miniprep J37016 which is currently in shaker since 11 am Wednesday
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