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<h2 align= "left">Test Overexpressions</h2> | <h2 align= "left">Test Overexpressions</h2> | ||
+ | <p class="pageText"> We conducted test overexpressions of our constructs in BL21 DE3 Gold, an <i>E. coli</i> strain capable of overexpressing T7 RNA polymerase by induction with IPTG. | ||
+ | </p> | ||
<h2 align= "left">Multi-Protein Purification</h2> | <h2 align= "left">Multi-Protein Purification</h2> |
Revision as of 22:52, 27 October 2017
Team is registered!
X parts characterized and documented
Provided simplified protocols and learning tools for the education system in collaboration with the Lethbridge High School Team
Project is showcased on wiki
Worked closely with user groups to inform our design
Addressed policy issues regarding cell-free systems
Project attributions clearly detailed
Assisted Lethbridge High School with lab work and received help with education interviews
Assessed the biosecurity implications of our project
Safety forms submitted
Collaborated with Florida State University (add details)
Developed and tested software tools for biocontainment and to mitigate dual-use
Judging form completed
Improved X parts by optimizing coding sequences for optimal expression in E. coli and for simple purification
Parts documented on the registry
Demonstrated proof of concept for multi-protein purification to simplify the system
X (list #) parts submitted
Participated in the InterLab study
TX-TL Proteins
Test Overexpressions
We conducted test overexpressions of our constructs in BL21 DE3 Gold, an E. coli strain capable of overexpressing T7 RNA polymerase by induction with IPTG.
Multi-Protein Purification
tRNA
tRNA Purification
Validation Construct
In vitro Transcription
To confirm our ability to detect successful transcription, the spinach aptamer was in vitro transcribed using T7 RNA polymerase (previously purified) and purified by phenol chloroform extraction. Following addition of the fluorophore, DFHBI, fluorescence was measured using a fluorimeter. The fluorimeter data illustrates that fluorescence was observed following addition of DFHBI, with an excitation of 447nm and emission of 497nm.