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??


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