Team:ETH Zurich/Biology/Journal

From 2011.igem.org

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Revision as of 15:55, 20 September 2011

Can you feel the smoke tonight?

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

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

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

Week 11: 29.8-4.9

Transformation and Ligation
- P_tet-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

Design of a system to characterize our new biobrick LacI_M1
- Transformation and Ligation
  - P_Tet and LacI
  - P_Lac and GFP assam

Design of an additional construct to test the band pass filter
- Transformation and Ligation
  - P_const and TetR_LVA

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
  - P_const and XylR
  - P_Xyl and CI
  - P_Xyl 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
- P_const-TetR- P_Tet-LacI_M1 and λP-GFPassam
Western Blot of AlcR
Making of competent DH5-α cells
Design of Psb6A5

Week 14: 19.9.-25.9

Characterization of LacI_M1

@@ Line 151: / Line 151: @@
 ***P<sub>const</sub> and TetR<sub>LVA</sub>
-*Growth-test of ''E.coli'' in M9 with reduced Ampicillin concentration
+*Growth-test of ''E. coli'' in M9 with reduced Ampicillin concentration
 *AlcR test with reduced Ampicillin concentration
@@ Line 165: / Line 165: @@
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 == '''Week 13:   12.9.-18.9''' ==