Calgary/full procedure
From 2007.igem.org
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Revision as of 17:46, 12 October 2007
EcoLisa is our Universities biological entry in the IGEM compeition. The goal of this project was to design a biomechanical printer. This printer works by using a laser to induce engineered E. coli produce agarase. The agarase will then dissolve the agar medium in which the E. coli is growing. The following procedure outlines the process our team went through to develop this project. A much simplified version of how our project works can be found in our Simplified Project Plan Section
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Choosing Our Project |
There were several different projects being considered by our team.
- Hydrogen Sulfide Sensor - This project involves designing cells that undergo an easily detectable change as a response to detecting H2S in the atmosphere.
- Bio Printer - Full description of this project is on... well the entire page.
- Google Cell - Create an addressable cell.
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Preparation Work |
Design primers to isolate agarase. Had to deal with issues of restriction site within the gene
Oderdering parts
Preparing Reagents
Preparing Plates. AMP resistant
Have everything ready to go for when parts arrive
Quick list of parts...
- R0084 - Light Sensor Promoter
- R0062 - AHL Promoter
- R0011 - Temperature Sensitive Promotor
- S03600 - AHL Intermediate
- I13544 - GFP RBS Terminator
- J23008 - RNA Key
- B0034 - RBS
- B0015 - Terminator
- J06501 - Temperature Sensitive Component
- I51001 - ccdb and AMP resistant (death gene used to do plasmid switches
Wells from registery
I) Rehydrate the wells
II) 2 micro litres of rehydrated part --> transformed into TOP10
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Testing The Parts From MIT |
I) Colony PCR test...
- Design universal primers (valid for any biobrick out of any plasmid)
- Develop massive amounts
- Gel to test
II) Test Plasmids --> always grow up 5ml of overnight
- Plasmid Prep --> isolating plasmid out of the cell
- Sigma Plasmid Prep protocol
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Constructing Our Parts |
Put designs
GFP as a test that will be replaced with agarase
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Lawn and Printer |
prepare bacterial broth then pour as agar.
Set up printer system
