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<h3>Introduction </h3> | <h3>Introduction </h3> | ||
− | <h5> We believed that supercoiling has been a problem for synthetic biologists trying to construct plasmids expressing multiple genes. We created plasmids to demonstrate supercoiling and the interference it creates between two genes. Our plasmids contain one GFP coding device and one RFP coding device, in varying orientations, with either a 500bp spacer, 1000bp spacer, or dCas9 clamp binding sites between them. The plasmids use the backbone pSB1C3 or <a href="https://www.addgene.org/66067/"> DVK_AE</a (derived from pSB1K3). Because the GFP and RFP devices are placed in different orientations we can observe whether or not supercoiling is present and whether or not it was reduced. (For more details, see <a href="https://2016.igem.org/Team:Alverno_CA/Design">Design</a>) After creating successful plasmids, we transformed them in E. coli. The colonies were then inoculated and grown as overnight liquid cultures. </p> | + | <h5> We believed that supercoiling has been a problem for synthetic biologists trying to construct plasmids expressing multiple genes. We created plasmids to demonstrate supercoiling and the interference it creates between two genes. Our plasmids contain one GFP coding device and one RFP coding device, in varying orientations, with either a 500bp spacer, 1000bp spacer, or dCas9 clamp binding sites between them. The plasmids use the backbone pSB1C3 or <a href="https://www.addgene.org/66067/"> DVK_AE</a> (derived from pSB1K3). Because the GFP and RFP devices are placed in different orientations we can observe whether or not supercoiling is present and whether or not it was reduced. (For more details, see <a href="https://2016.igem.org/Team:Alverno_CA/Design">Design</a>) After creating successful plasmids, we transformed them in E. coli. The colonies were then inoculated and grown as overnight liquid cultures. </p> |
<h3>Experiment </h3> | <h3>Experiment </h3> |
Revision as of 21:25, 19 October 2016