IGEM:MIT/2007/Notebook/2007-7-31

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Agenda

  1. Run mercury assay
  2. Redo live immunostaining with altered protocol
  3. Enter working parts into BioBricks Registry
  4. Find constitutive promoter and perform 3A assembly with a RBS and CPX
  5. Redo transformation of BL21 w/ mer.I13500 in 1AT3

Mercury Assay

CPX Assay

Varied cell concentrations: (OD)

  • .1
  • .3
  • .5
  • 1

CPX Immunostaining

  • Induced cells at 0.1 OD, incubate for 1 hour
    • 10 samples total: 2 each for 0(- control), 10^-8, 10^-7, 10^-6, 10^-5 concentration AHL
  • Changed secondary Goat Anti-Mouse antibody concentration to (1:200)

Results

  • AHL 5Ai

    AHL 5Ai

  • AHL 5Aii

    AHL 5Aii

  • AHL 5Bi

    AHL 5Bi

  • AHL 5Bii

    AHL 5Bii

  • AHL 6Ai

    AHL 6Ai

  • AHL 6Aii

    AHL 6Aii

  • AHL 6Bi

    AHL 6Bi

  • AHL 7Ai

    AHL 7Ai

  • AHL 7Aii

    AHL 7Aii

  • AHL 7Aiii

    AHL 7Aiii

  • AHL 7Bi

    AHL 7Bi

  • AHL 7Bii

    AHL 7Bii

  • AHL 8Ai

    AHL 8Ai

  • AHL 8Aii

    AHL 8Aii

  • AHL 8Bi

    AHL 8Bi

  • AHL 8Bii

    AHL 8Bii

  • Control Ai

    Control Ai

  • Control Aii

    Control Aii

  • Control Bi

    Control Bi

  • Control Bii

    Control Bii

Transformation of mer.I13500 in BL21

  • Nothing grew on any of the plates (not even positive control with pUC18)
  • Perhaps cells not competent?
  • We should redo tomorrow
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