IGEM:IMPERIAL/2007/Projects/Experimental Design
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We have identified three phases of experiments for each of the project:
- Phase 1 - Intial testing of the promoters and test constructs in vitro
- Phase 2 - Testing/Validation of the test constructs
- Phase 3 - Testing/Validation of final constructs
In addition any overlap between the experiments of the projects has been identified and considered when making the experimental design.
Status
- Several Problems have been identified, we have begun to address them. Link here to problems.
- We are currently trying to improve our methodology to get better results. Link to improving our Methodology
- Problems with dsRED, it has failed quality control
- Currently trying to purify LuxR and GFP.
Time table
General timetable
Week 8 | Week 9 | Week 10 | |
Monday | - | Refine the wiki | Planning the week |
Tuesday | Prepare GFP solution-calibration Construct Calibration Curve Refine the wiki |
Testing to reduce evaporation | |
Wednesday | Test Home made cell extract Test varying levels of cell extract |
PROBLEM-Wrong GFP!!!! | 24hr experiment with pTet using paraffin oil |
Thursday | |||
Friday | Prepare GFP solution-calibration Construct Calibration Curve |
CBD
Week 8 | Week 9 | Week 10 | |
Monday | - | ||
Tuesday | Phase 1 - test pT7 | ||
Wednesday | |||
Thursday | Test 20 and 37 °C, Test GFP degradation at 20 and 37 °C |
24 Hour Exp Room Temp | |
Friday | Second day testing 20 and 37 °C | Define the thresholds of temperature |
ID
Week 8 | Week 9 | Week 10 | |
Monday | - | Purify LuxR | |
Tuesday | test pLux in-vitro - 1nM AHL | Refine AHL concentrations range 25°C Purify LuxR if plasmid arrives |
Purification of LuxR - problems |
Wednesday | Purify LuxR if plasmid arrives |
problems with LuxR pufification | |
Thursday | Clone in plasmid | ||
Friday | Test construct with 5 AHL concentrations - 25 °C | Begin purification of LuxR | Test Purified LuxR with construct 2?? |