Team:Wash U/Protocol
From 2009.igem.org
(Difference between revisions)
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:1. Begin BioBrick Assembly with three separate plasmids, an upstream part, a down stream part, and a destination plasmid. It is important that the destination plasmid contain the toxic gene ccdB in the BioBrick cloning site and a different antibiotic resistance to the upstream and downstream parts.<br> | :1. Begin BioBrick Assembly with three separate plasmids, an upstream part, a down stream part, and a destination plasmid. It is important that the destination plasmid contain the toxic gene ccdB in the BioBrick cloning site and a different antibiotic resistance to the upstream and downstream parts.<br> | ||
:2. Digest each part with the proper restriction enzymes to isolate the upstream and downstream parts, and to remove the BioBrick from the destination plasmid.<br> | :2. Digest each part with the proper restriction enzymes to isolate the upstream and downstream parts, and to remove the BioBrick from the destination plasmid.<br> | ||
- | :3. | + | :3. Ligate the products. The result will be the upstream part connected to the downstream part in the destination plasmid. This new composite part can then be used to transform competent cells.<br> |
:To view the full Biobrick Assembly manual, click [http://ginkgobioworks.com/support/ here]. | :To view the full Biobrick Assembly manual, click [http://ginkgobioworks.com/support/ here]. | ||
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- | + | '''Ligation''' | |
:text | :text | ||
Revision as of 17:06, 30 June 2009