IGEM:Arizona State/2014/Notebook/asu igem 2014/2014/07/08: Difference between revisions
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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span> | |style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span> | ||
|style="background-color: #F2F2F2" align="center"| | |style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]] }}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}} | ||
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Latest revision as of 00:05, 27 September 2017
Project name | Main project page Previous entry Next entry |
Gel Extraction, New Digests, and Tentative Plans
Digest RBS at S and P (at 3pm) and run overnight. Already have Tes cut at X and P. Later Gel: Run in gel and extract TesA part without backbone and RBS linear plasmid with backbone. Ligate: TesA part into RBS backbone part E|| RBS | S| X| TESA | P|| P (ready to cut at ends) Transform: Transform the plasmids into cells (mix plasmids with competent cells) Both plasmids into cell Plate Cells Pick colonies to make seeds/tubes Miniprep seed cultures, and gel extract with complete plasmid Digest P and S sites at end of Tes, and Ligate MCherry at X and P sites Gel extract longest piece (all together RBS, TES, and MCherry) E| RBS | S| X| TESA | S| X| MCherry | P Dr. Nielsons Further Suggestions (in case this procedure doesn't work) 1. Two plasmids 2. Attach primers to TesA and MC 3. Order Tes with RBS attached 4. Start ACL with Wax 5. Might not add MCherry to plasmid, instead add to chromosome to make permanent change later. 6. Use PCR to complete some of steps
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