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From 2011.igem.org

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 for the system
• Design of several Operons for AlcR and several PCR primer

Week 4: 11.7-17.7

• Ordering of the LacI_M1 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
  • λ_P and luxI
  • λ_P and lacI
  • P_lac and GFP_LVA
  • P_lux and mCherry
  • P_tet and CI -had to be redone because of point mutation in the primer (week 9)
  • P_const 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

• Synthesized LacI_M1 Gen arrived

• Ligation and Transformation of
  • P_lac-GFP_LVA and Terminator
  • P_lux-mCherry and Terminator
  • P_const-luxR and Terminator
  • P_lac-GFP_LVA-Terminator and P_lux-mCherry-Terminator on one plasmid
  • P_tet and LacI_M1
  • P_tet-LacI_M1 and Terminator

• Improving of the testsystem
  • Exchange of same rare codons in AlcR by PCR
  • His-tagging of AlcR
  • GFP assam

• Transformation of λ_P

• 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
  • λ_P and lacI
  • λ_P and luxI
  • P_tet and CI
  • λ_P-lacI and terminator
  • λ_P-luxI and terminator

Week 10: 22.8-28.8