IGEM:IMPERIAL/2006/LabCalendar/2006-8-18

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Contents

Maxipreps

  • 12D - for J37020 ligation
  • 5M - for ligating into aiiA construct when primers arrive
  • J37016 - so a sample can be sent for sequencing

Sequencing

  • J37016 sent for sequencing
    • 1 μg DNA
    • 12.5 pmol I13504 primer (CAA CTA GCA GAC CAT TAT C)

Minipreps

  • J37015RS
    • We have innoculated a 100 mL culture in the fridge to be maxipreped on Monday
    • All cultures were successful; however, we took only colony 4 since it showed the best looking bands on the gel (yes, I guess we do have to be discriminatory)
  • 1M
    • Band at around 2000 showing the plasmid, the part itself is too small and may have run too far, so we assume that it is correct
    • This part can be maxipreped on Monday along with J37015RS
  • 4G
    • Successful miniprep, we can start testing this on Monday as it is an entire part ready to be testing. Please ensure that you culture in Kanamycin LB (located in fridge)
  • 2H
    • Unsuccessful, needs redoing
    • Second miniprep unsuccessful. Have to let MIT know we have been struggling to ligate part 2H and that we suspect that it may involve a mutation in the Spe1 site.


Ligations

  • J37031 = J37019 + B0015 (1I)
  • J37032 = R0062 (9G) + I13504 (12D)
    • 12D on the gel after digestion was not clean, this might pose a small problem, but I cut out the smaller band anyway that look around 800 bp (compared to the J37019 which is also about 800 bp).
  • Electroporations this afternoon
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