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


 * Running gel of yesterday's gradient PCR:
 * 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: Failed:
 * I2057-3K3
 * T9002-3K3 and 1AK3
 * E0240-3K3
 * 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.