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Team:Calgary/Notebook/PostRegionals
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<h2>Biotinylation - <a href="https://2011.igem.org/Team:Calgary/Project/ProjectPseudomonas#biotin">Click Here For More</a> | <h2>Biotinylation - <a href="https://2011.igem.org/Team:Calgary/Project/ProjectPseudomonas#biotin">Click Here For More</a> | ||
</h2><ul> | </h2><ul> | ||
| - | <li>Successfully | + | <li>Successfully biotinylated two new naphthenic acids (cyclohexanebutyric acid and cyclohexanecarboxylic acid) identifying the ability of our biotinylation method to be used with compounds of varying hydrophobicity.</li> |
| - | <li>Characterized the binding of our biotin linker to a variety of types | + | <li>Characterized the binding of our biotin linker to a variety of bead types and determined a protocol by which biotinylated hydrophobic compounds can bind to Invitrogen Dynabeads.</li> |
<li>Developed and ran an immunoprecipitation protocol providing evidence for the "pull-down" of proteins which bind CHPA. </li></ul> | <li>Developed and ran an immunoprecipitation protocol providing evidence for the "pull-down" of proteins which bind CHPA. </li></ul> | ||
| Line 28: | Line 28: | ||
<h2>Conjugation - <a href="https://2011.igem.org/Team:Calgary/Project/Chassis/Pseudomonas#conresults">Click Here For More</a></h2> | <h2>Conjugation - <a href="https://2011.igem.org/Team:Calgary/Project/Chassis/Pseudomonas#conresults">Click Here For More</a></h2> | ||
<ul> | <ul> | ||
| - | <li>Characterized our ori1600/oriT conjugation system against Berkeley's oriT submitted in 2005 ( | + | <li>Characterized our ori1600/oriT conjugation system against Berkeley's oriT submitted in 2005 (BBa_J01003), quantifying the relative rate of transformation. This data suggested our oriT construct has a higher efficiency of conjugation than Bba_J01003.</li> |
<li>Optimized protocol for single colonies, and demonstrated the robustness of this conjugation system.</li></ul> | <li>Optimized protocol for single colonies, and demonstrated the robustness of this conjugation system.</li></ul> | ||
Latest revision as of 04:03, 29 October 2011
Notebook
Post-Regionals
Journal
- Defining Objectives
- Techniques Workshop
- Bioinformatics & qRT-PCR
- Sensory Element Fishing
- Electrochem - CPR Oxidation
- Electrochem - Reporter Gene
- Pseudomonas Tools
- Microalgae Tools
- NA Viability Assays
- May - July Group Journal
Protocols
Safety
Post-Regionals
The America's Regional Competition
The University of Calgary iGEM Team had great success at the Regional Jamborree held in Indianapolis, Indiana this year, winning the Best Wiki and Best Measurement special prizes, obtaining a gold medal, and advancing as a finalist to the world's Jamborree at MIT. In the few weeks between competitions, our team has been hard at work to clarify, optimize, and rework aspects of our project. Here is a list of our major accomplishments in the last three weeks:
Prototype - Click Here For More
- Developed a prototype system composed of a portable potentiostat capable of making readings using our lacZ system
- Produced a composite part Bba_K640006 composed of a response element we identified from the fad-2 gene in Pseudomonas fluorescens, the lacZ part, oriT, and ori1600, making it potentially a completely functional circuit which could be used to measure naphthenic acids in solution. (Further characterization is required, however, and is being pursued by the team up until the competition)
Biotinylation - Click Here For More
- Successfully biotinylated two new naphthenic acids (cyclohexanebutyric acid and cyclohexanecarboxylic acid) identifying the ability of our biotinylation method to be used with compounds of varying hydrophobicity.
- Characterized the binding of our biotin linker to a variety of bead types and determined a protocol by which biotinylated hydrophobic compounds can bind to Invitrogen Dynabeads.
- Developed and ran an immunoprecipitation protocol providing evidence for the "pull-down" of proteins which bind CHPA.
LacZ Electrochemical Detector - Click Here For More
- Optimized the electrochemical buffer system and electroplating methods to gain an electrochemical detection system operable at high effiency in field conditions.
Conjugation - Click Here For More
- Characterized our ori1600/oriT conjugation system against Berkeley's oriT submitted in 2005 (BBa_J01003), quantifying the relative rate of transformation. This data suggested our oriT construct has a higher efficiency of conjugation than Bba_J01003.
- Optimized protocol for single colonies, and demonstrated the robustness of this conjugation system.
Naphthenic Acid BioSensor Marketing Analysis - Click Here For More
- In order to show the feasibility of using a naphthenic acid biosensor in a real world application, our team has prepared a report identifying a market for our biosensor, a comprehensive cost analysis of our product, and a comparison of our product to traditional and other biosensor methods already available in the market.
Microalgae Parts - Click Here For More
- Developed a testing construct for the algae optimized luciferase part.
