Team:ETH Zurich/Biology/Journal
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 of the system
- 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
- 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