Team:ETH Zurich/Biology/Journal

From 2011.igem.org

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*Transformation and Ligation of
*Transformation and Ligation of
** 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
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*Western Blot of AlcR
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* Western Blot of AlcR
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*Making of competent DH5-α cells  
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* Making of competent DH5-α cells  
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* Design of Psb6A5
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Revision as of 14:30, 19 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
  • Making of competent DH5-α cells
  • Design of Psb6A5

Week 14: 19.9.-25.9

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