Registry/Measurement kit/Notebook/2007-7-11

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  • Running gel of yesterday's gradient PCR:

<order?>

  • Getting strange secondary product on E0240 BB pcr. Should re-try with vent polymerase.
  • Did so (but ran out of template dna in the process):

Sequencing results

  • Newly constructed I2055 doesn't have promoter in it
  • E0240-1AK3 looks scarred from reverse. Forward sequence is too short; trying again with primers internal to GFP.
  • T9002-3K3 just matched 3K3


Ligation

Growth on:

  • I2057-3K3
  • T9002-3K3 and 1AK3
  • E0240-3K3

Failed:

  • I2057-1AK3 (strange growth)
  • I2056-1AC3, 1AT3 (none)
  • Re-plating these

Colony PCR

  • Row 1: I2057-3K3 (4-1) [1], I2057-1AK3(1-4) E0240-3K3(1-4) [1]
  • Row 2: T9002-3K3 (4-1) [3], T9002-1AK3 [3]
  • brackets indicate colony chosen for sequencing
  • 3K3 may be working, but mis-priming in the PCR. Will check with internal GFP primers.
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