IGEM:MIT/2007/Notebook/2007-7-31: Difference between revisions

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==Agenda==
=Agenda=
#Run mercury assay
#Run mercury assay
#Redo live immunostaining with altered protocol
#Redo live immunostaining with altered protocol
Line 6: Line 6:
#Redo transformation of BL21 w/ mer.I13500 in 1AT3
#Redo transformation of BL21 w/ mer.I13500 in 1AT3


==Mercury Assay==


==Results of CPX 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====
<gallery>
<gallery>
Image:AHL-6 1.jpg|AHL-6 1
Image:8-1-AHL-5Ai.jpg|AHL 5Ai
Image:AHL-6 2.jpg|AHL-6 2
Image:8-1-AHL-5Aii.jpg|AHL 5Aii
Image:AHL-6 3.jpg|AHL-6 3
Image:8-1-AHL-5Bi.jpg|AHL 5Bi
Image:AHL -7 1.jpg|AHL-7 1
Image:8-1-AHL-5Bii.jpg|AHL 5Bii
Image:AHL -7 2.jpg|AHL-7 2
Image:8-1-AHL-6Ai.jpg|AHL 6Ai
Image:AHL-7 3.jpg|AHL-7 3
Image:8-1-AHL-6Aii.jpg|AHL 6Aii
Image:AHL-8 1.jpg|AHL-8 1
Image:8-1-AHL-6Bi.jpg|AHL 6Bi
Image:AHL-8 2.jpg|AHL-8 2
Image:8-1-AHL-7Ai.jpg|AHL 7Ai
Image:AHL-8 3.jpg|AHL-8 3
Image:8-1-AHL-7Aii.jpg|AHL 7Aii
Image:Control 1.jpg|Control 1
Image:8-1-AHL-7Aiii.jpg|AHL 7Aiii
Image:Control 2.jpg|Control 2
Image:8-1-AHL-7Bi.jpg|AHL 7Bi
Image:Control 3.jpg|Control 3
Image:8-1-AHL-7Bii.jpg|AHL 7Bii
Image:8-1-AHL-8Ai.jpg|AHL 8Ai
Image:8-1-AHL-8Aii.jpg|AHL 8Aii
Image:8-1-AHL-8Bi.jpg|AHL 8Bi
Image:8-1-AHL-8Bii.jpg|AHL 8Bii
Image:8-1-controlAi.jpg|Control Ai
Image:8-1-controlAii.jpg|Control Aii
Image:8-1-controlBi.jpg|Control Bi
Image:8-1-controlBii.jpg|Control Bii
</gallery>
</gallery>
==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

Latest revision as of 14:11, 2 August 2007

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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