Team:ETH Zurich/Biology/Journal

From 2011.igem.org

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|colspan="2"|'''Here you can follow the progress of our project as it is coming together.'''
|colspan="2"|'''Here you can follow the progress of our project as it is coming together.'''
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== '''Week 1:  20.6-26.6''' ==
== '''Week 1:  20.6-26.6''' ==
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* Brainstorming
* Brainstorming
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== ''' Week 2:  27.6-3.7''' ==
== ''' Week 2:  27.6-3.7''' ==
* Brainstorming
* Brainstorming
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== ''' Week 3:  4.7-10.7''' ==
== ''' Week 3:  4.7-10.7''' ==
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* Design of several Operons for AlcR and several PCR primer
* Design of several Operons for AlcR and several PCR primer
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== '''Week 4:  11.7-17.7''' ==
== '''Week 4:  11.7-17.7''' ==
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* Making of competent DH5-α cells
* Making of competent DH5-α cells
* Cloning of the AlcR-testsystem
* Cloning of the AlcR-testsystem
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== '''Week 5:  18.7-24.7''' ==
== '''Week 5:  18.7-24.7''' ==
* First test of the AlcR-system in 96-well plates
* First test of the AlcR-system in 96-well plates
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== '''Week 6:  25.7-31.7''' ==
== '''Week 6:  25.7-31.7''' ==
* Transformation of the parts from the iGEM plates
* Transformation of the parts from the iGEM plates
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== '''Week 7:  1.8-7.8''' ==
== '''Week 7:  1.8-7.8''' ==
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* First meeting with Dr. Oliver Frey (Bio engineering laboratory, Prof. Andreas Hierlemann, D-BSSE ETHZ) to exchange ideas about microfluidic design
* First meeting with Dr. Oliver Frey (Bio engineering laboratory, Prof. Andreas Hierlemann, D-BSSE ETHZ) to exchange ideas about microfluidic design
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== '''Week 8:  8.8-14.8''' ==
== '''Week 8:  8.8-14.8''' ==
[[File:Lab5.jpg|right|300px]]
[[File:Lab5.jpg|right|300px]]
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* Making of competent JM101 cells
* Making of competent JM101 cells
** Check for transformation efficiency
** Check for transformation efficiency
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== '''Week 9:  15.8-21.8''' ==
== '''Week 9:  15.8-21.8''' ==
* Growth and repression test of AlcR-system with M9-medium
* Growth and repression test of AlcR-system with M9-medium
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** &lambda;<sub>P</sub>-lacI and terminator
** &lambda;<sub>P</sub>-lacI and terminator
** &lambda;<sub>P</sub>-luxI and terminator
** &lambda;<sub>P</sub>-luxI and terminator
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== '''Week 10:  22.8-28.8''' ==
== '''Week 10:  22.8-28.8''' ==
* Transformation and Ligation
* Transformation and Ligation
** P<sub>tet</sub>-CI and Terminator
** P<sub>tet</sub>-CI and Terminator
*Preparation of chemically competent DH5-α and JM101 cells
*Preparation of chemically competent DH5-α and JM101 cells
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== '''Week 11:  29.8-4.9''' ==
== '''Week 11:  29.8-4.9''' ==
* Transformation and Ligation
* Transformation and Ligation
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* Diffusion test through tube system filled with agarose and cells
* Diffusion test through tube system filled with agarose and cells
* Test of the improved AlcR system
* Test of the improved AlcR system
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== '''Week 12:  5.9.-11.9''' ==
== '''Week 12:  5.9.-11.9''' ==
[[File:Lab6.JPG|right|300px]]
[[File:Lab6.JPG|right|300px]]
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*** P<sub>Xyl</sub> and CI
*** P<sub>Xyl</sub> and CI
*** P<sub>Xyl</sub> and LacI <sub>M1</sub>
*** P<sub>Xyl</sub> and LacI <sub>M1</sub>
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== '''Week 13:  12.9.-18.9''' ==
== '''Week 13:  12.9.-18.9''' ==
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** P<sub>const</sub>-TetR- P<sub>Tet</sub>-LacI<sub>M1</sub> and  λP-GFPassam
** P<sub>const</sub>-TetR- P<sub>Tet</sub>-LacI<sub>M1</sub> and  λP-GFPassam
*Western Blot of AlcR
*Western Blot of AlcR
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== '''Week 14:  19.9.-25.9''' ==
== '''Week 14:  19.9.-25.9''' ==
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Revision as of 15:27, 15 September 2011

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Lab Journal
Week 1 2 3 4 5 6 7 8 9 10 11 12 13 14
Here you can follow the progress of our project as it is coming together.

Week 1: 20.6-26.6

  • First meeting
  • Brainstorming

Week 2: 27.6-3.7

  • Brainstorming

Week 3: 4.7-10.7

  • Working on the design of the system
IMG 5009.JPG
IMG 5010.jpg
  • Design of several Operons for AlcR and several PCR primer

Week 4: 11.7-17.7

  • Ordering of the LacIM1 and codonoptimized AlcR gene
  • Making of competent DH5-α cells
  • Cloning of the AlcR-testsystem

Week 5: 18.7-24.7

  • First test of the AlcR-system in 96-well plates

Week 6: 25.7-31.7

  • Transformation of the parts from the iGEM plates

Week 7: 1.8-7.8

  • Growth test of DH5-α with AlcR-system in flasks
  • Ligation and Transformation of
    • λP and luxI
    • λP and lacI
    • Plac and GFPLVA
    • Plux and mCherry
    • Ptet and CI -had to be redone because of point mutation in the primer (week 9)
    • Pconst and luxR
  • First meeting with Dr. Oliver Frey (Bio engineering laboratory, Prof. Andreas Hierlemann, D-BSSE ETHZ) to exchange ideas about microfluidic design

Week 8: 8.8-14.8

Lab5.jpg
  • Synthesized LacIM1 Gen arrived
  • Ligation and Transformation of
    • Plac-GFPLVA and Terminator
    • Plux-mCherry and Terminator
    • Pconst-luxR and Terminator
    • Plac-GFPLVA-Terminator and Plux-mCherry-Terminator on one plasmid
    • Ptet and LacIM1
    • Ptet-LacIM1 and Terminator
  • Improving of the testsystem
    • Exchange of same rare codons in AlcR by PCR
    • His-tagging of AlcR
    • GFP assam
  • Transformation of λP
  • Making of competent JM101 cells
    • Check for transformation efficiency

Week 9: 15.8-21.8

  • Growth and repression test of AlcR-system with M9-medium
  • Ligation and Transformation of
    • λP and lacI
    • λP and luxI
    • Ptet and CI
    • λP-lacI and terminator
    • λP-luxI and terminator

Week 10: 22.8-28.8

  • Transformation and Ligation
    • Ptet-CI and Terminator
  • Preparation of chemically competent DH5-α and JM101 cells

Week 11: 29.8-4.9

  • Transformation and Ligation
    • Ptet-CI and Terminator
  • Diffusion test through tube system filled with agarose and cells
  • Test of the improved AlcR system

Week 12: 5.9.-11.9

Lab6.JPG
  • Design of a system to characterize our new biobrick LacIM1
    • Transformation and Ligation
      • PTet and LacI
      • PLac and GFP assam
  • Design of an additional construct to test the band pass filter
    • Transformation and Ligation
      • Pconst and TetRLVA
  • Growth-test of E.coli in M9 with reduced Ampicillin concentration
  • AlcR test with reduced Ampicillin concentration
  • Testdigest of λP-luxI-Terminator
  • Alternative Sensor system: Xylene system
    • Transformation and Ligation
      • Pconst and XylR
      • PXyl and CI
      • PXyl and LacI M1

Week 13: 12.9.-18.9

  • PCR of the pBR323 Ori
  • PCR of the gen cluster for the upper TOL pathway
  • first test of cell printing
  • Transformation and Ligation of
    • Pconst-TetR- PTet-LacIM1 and λP-GFPassam
  • Western Blot of AlcR

Week 14: 19.9.-25.9

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