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Team:BostonU/Chimera
From 2014.igem.org
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<th scope="col">The lack of standardization. this will become an increasingly significant stymying factor to the growth of the field as more laboratories continue to share resources and data. We seek to strengthen the traditional design-build-test cycle fundamental to synthetic biology with a formalized workflow defined by bio-design automation software tools and built upon a thoroughly characterized library of parts.</th> | <th scope="col">The lack of standardization. this will become an increasingly significant stymying factor to the growth of the field as more laboratories continue to share resources and data. We seek to strengthen the traditional design-build-test cycle fundamental to synthetic biology with a formalized workflow defined by bio-design automation software tools and built upon a thoroughly characterized library of parts.</th> | ||
| - | <th scope="col"><img src="https://static.igem.org/mediawiki/2014/d/d9/Chimera_plasmid_BU14.png" alt="ChimeraPlasmid" style="float:right" style= "margin-left:10px"><br><br><capt><br>THE CHIMERA</capt></th> | + | <th scope="col"><img src="https://static.igem.org/mediawiki/2014/d/d9/Chimera_plasmid_BU14.png" height="300" width="300" alt="ChimeraPlasmid" style="float:right" style= "margin-left:10px"><br><br><capt><br>THE CHIMERA</capt></th> |
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Revision as of 20:04, 11 July 2014
| The lack of standardization. this will become an increasingly significant stymying factor to the growth of the field as more laboratories continue to share resources and data. We seek to strengthen the traditional design-build-test cycle fundamental to synthetic biology with a formalized workflow defined by bio-design automation software tools and built upon a thoroughly characterized library of parts. |  THE CHIMERA |
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June |
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Week of June 23 |
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| The backbones that would have their origin replaced were selected and new origins were selected. DVL1 with "A" and "E" MoClo fusion sites and DVL2 with "A" and "F" fusion sites were initially chosen, as they are the most commonly used MoClo level 1 and 2 destination vectors, respectively (See MoClo for more information on our assembly method). The general plan to replace the backbones was formulated, which comprised of: 1. Using PCR to extract the backbones without their high-copy origins from their full destination vectors, and to extract the lower-copy origins from their respective plasmids. 2.Performing a restriction digest on the backbone and origin fragments to have compatible sticky ends. 3. Ligating the origins to the destination vectors. 4. Transforming into E. Coli, purifying the plasmid DNA, and sequencing for confirmation. The PCR primer design added restriction sites for the MfeI restriction enzyme, which would give the ends of each of the amplified fragments compatible 4bp overhangs suitable for ligation. (Detailed primer design available here). • Struck out devices with low copy origins for PCR on plates with appropriate antibiotic. • Prepared liquid cultures, incubated, and miniprepped. • Received primers, diluted. |
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