IGEM:IMPERIAL/2007/Projects/Experimental Design

• Introduction
• Phase 1
• Phase 2
• Phase 3
• Notes

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.

Time table

Testing of constructs and cell extracts

	Week 8	Week 9	Week 10
Monday	-	Refine the wiki
Tuesday		Prepare GFP solution-calibration Construct Calibration Curve Refine the wiki
Wednesday	Test Home made cell extract Test varying levels of cell extract
Thursday
Friday	Prepare GFP solution-calibration Construct Calibration Curve

CBD

	Week 8	Week 9	Week 10
Monday	-		Exp with 15 and 25 °C
Tuesday	Phase 1 - test pT7		Exp with 30 and 40 °C, continue measuring 15 and 25 °C
Wednesday			Exp with 10 and 45 °C, continue measuring 30 and 40 °C
Thursday	Test 20 and 37 °C, Test GFP degradation at 20 and 37 °C		Test suitable step increases
Friday	Second day testing 20 and 37 °C		Test suitable gradient of temperature, minimum working temp - maximum working temp

ID

	Week 8	Week 9	Week 10
Monday	-
Tuesday	test pLux in-vitro - 1nM AHL	Refine AHL concentrations range 25°C Purify LuxR if plasmid arrives
Wednesday		Choose a range of AHL concentrations range 37°C Purify LuxR if plasmid arrives
Thursday		Test purified LuxR
Friday	Test construct with 5 AHL concentrations - 25 °C	Test purified LuxR