IGEM:MIT/2007/Notebook/2007-8-9

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ACCOMPLISHED TODAY

Miniprepped Standard Assembly LCs

  • 12 tubes, 4 each of CPX/B0014, I14032/B0034 and R0051/B0034, all in DH5alpha
  • NOTE: Elution with Tris EDTA, not water
  • Nanodrop results obtained
CPX 1 : 103.3 ng/microliter
CPX 2 : 131.7
CPX 3 : 135.5
CPX 4 : 135.3

R0051 1 : 266.1
R0051 2 : 211
R0051 3 : 266.9
R0051 4 : 54.7

I14032 1 : 219.7
I14032 2 : 247
I14032 3 : 211.1
I14032 4 : 225.7

Diluted and Grew Up CPX-BL21 and Untransformed BL21 (for Western Blot on 8/10)

  • Induced for 1 hour with AHL
  • OD of samples at about 3.30 PM:
Untransformed BL21
[AHL] = 0; 1.24 OD; "U (-)"
[AHL] = 1E-8; 1.21 OD; "U (-8)"
[AHL] = 1E-5; 1.22 OD; "U (-5)"

CPX BL21
[AHL] = 0; 0.87 OD; "T (-)"
[AHL] = 1E-8; 0.41 OD; "T (-8)"
[AHL] = 1E-7; 0.36 OD; "T (-7)"
[AHL] = 1E-5; 0.35 OD; "T (-5)"
  • Took enough of each sample to be about 0.4 OD and spun down; placed in -20 degree freezer
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