Team:ETH Zurich/Biology/Journal

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Can you feel the smoke tonight?
 

Contents

Lab Journal

Here you can follow the progress of our project as it is coming together.

Week 1: 20.6-26.6

  • First meeting


Week 2: 27.6-3.7


Week 3: 4.7-10.7

  • Working on the design of the system
  • Design of several Operons for AlcR and several PCR primers


Week 4: 11.7-17.7

  • Ordering of the LacIM1 and codon-optimized 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
    • PR and luxI
    • PR 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 some rare codons in AlcR by PCR
    • His-tagging of AlcR for detection in Western blot
    • include sfGFP
  • Transformation of PR
  • Creation of competent JM101 cells
    • Check for transformation efficiency


Week 9: 15.8-21.8

IMG 6171 Kopie.JPG
  • Growth and repression test of AlcR-system with M9-medium
  • Ligation and Transformation of
    • PR and lacI
    • PR and luxI
    • Ptet and CI
    • PR-lacI and terminator
    • PR-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

IMG 6442.JPG
  • PCR of the pBR322 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 PR-GFPassam
  • Western Blot of AlcR
  • Making of competent DH5-α cells
  • Design and construction of Psb6A5


Week 14: 19.9.-25.9

  • Characterization of LacIM1
  • Cloning of all biobricks into pSB1C3
  • PCR of LacI and CI
  • wiki


Week 15: 26.9.-02.10

  • design of Pu promoter
  • test of Pu promoter
  • test for indol degradation
  • Poster
  • Preparing the presentation


European Jamboree 30.9-2.10

  • Sightseeing and having fun in Amsterdam
  • Gold Medal
  • Honorable Mention in category Best Model
  • Jamboree


Week 16: 03.10.-09.10

  • proof of concept with an arabinose inducable system
  • cloning for the xylene system
  • cloning of the lac-testsystem to compaire LacIM1


Week 17: 10.10.-16.10

  • cloning of the alarm-test system
  • cloning the final system with xylene
  • cloning the arabinose system
  • cloning the AlcR (codon optimized) system
  • building a channel out of different materials
  • Design a assay for testing the alarm system
  • Sender receiver test on plates


Week 18: 17.10.-23.10

  • Cloning
  • inactivating of the kanamycin casette of BW27749 to get BW27783
  • Make competend BW27783 and DH5-α cells
  • Sender receiver test on plates with sterile filtrated supernatend
  • Sender an receiver test in tube
  • Dose responce experiment for xylene
  • Dose responce experiment for acetaldehyde
  • Making a biobrick out of AraBAD (site directed mutatgenesis to remove restriction enzyme site)


Week 19: 24.10-30.10


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