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

• Running gel of yesterday's gradient PCR:

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• 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.