IGEM:IMPERIAL/2007/Projects/Cell by date/Implementation

Cell by Date: Implementation

• Introduction
• Specifications
• Design
• Modelling
• Implementation
• Testing/Validation
• Notes
• References

List of Required Protocols

Preliminary Preparation

• Endy:Making a long term stock of bacteria
• Endy:Retrieving a Registry glycerol

Phase 1

Chassis

Chassis	Level 1	Level 2	Level 3
E.coli	X
In-Vitro	X	X	X
In-Veso	X	X	X

Things to consider:

• Different types of promoters in different chassis
• Different threshold
• Different degradation time
• Different reporters

Preliminery Data

Protocols Required:

• Knight:Electroporation
• Endy:High-efficiency electroporation
• Incubation

1. Degradation time

• Endy:RNA Half-life
• λ-Reporters

2. Rate of expression at constant temperature

• Fluorometer Reading
• Noting down colour change (photographs)

3. Range of Reporters

• Flurometer Reading
  • RFP
  • DsRed

• Beta-Galactosidase_Assay_(A_better_Miller)
  • β-galactoside - requires subtrates X-GAL

• Luciferase - requires substrate ATP, O₂, and luciferin

4. Range of promoters

Refer to List of Required Parts.

Data Analysis

Refer to Modelling Tab.

Validation of Model

1. Validate [Reporter]_max and V_max value

• Fluorometer Reading
• Noting down colour change (photographs)

2. Validate transient response using best promoters/reporters

• Fluorometer Reading
• Noting down colour change (photographs)