Team:UC Davis/notebook/initiator.html
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<table class="pikachu" width="690px"> | <table class="pikachu" width="690px"> | ||
<tr><td class="kirby"> | <tr><td class="kirby"> | ||
- | <p class="header"><b>The Initiator</b></a><p> | + | <p class="header"><b>Assembly Workplan: The Initiator</b></a><p> |
+ | |||
+ | <img src="https://static.igem.org/mediawiki/2010/2/29/Initiatorconstruct.jpg"><p> | ||
+ | |||
+ | Because the initiator is one of the smaller constructs, we have decided to use this as an example to illustrate the utmost basic workplan in greater detail. <p> | ||
<a name="startup"><h1>Starting Up</h1></a> | <a name="startup"><h1>Starting Up</h1></a> | ||
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</ul><p><br /> | </ul><p><br /> | ||
- | <a name="rbsc0051"><h1> | + | <a name="rbsc0051"><h1>RBS to BBa_C0051</h1></a> |
<ul> | <ul> | ||
- | <li><a href="https://2010.igem.org/Team:UC_Davis/protocols/culture.html" class="help">Cultured</a> cells | + | <li><a href="https://2010.igem.org/Team:UC_Davis/protocols/culture.html" class="help">Cultured</a> cells that have parts: BBa_B0034 and BBa_C0051</li> |
<li><a href="https://2010.igem.org/Team:UC_Davis/protocols/miniprep.html" class="help">Miniprepped</a> BBa_B0034 and BBa_C0051</li> | <li><a href="https://2010.igem.org/Team:UC_Davis/protocols/miniprep.html" class="help">Miniprepped</a> BBa_B0034 and BBa_C0051</li> | ||
</ul> | </ul> | ||
Line 52: | Line 56: | ||
<li><a href="https://2010.igem.org/Team:UC_Davis/protocols/gelelectrophoresis.html" class="help">Run on gel</a></li> | <li><a href="https://2010.igem.org/Team:UC_Davis/protocols/gelelectrophoresis.html" class="help">Run on gel</a></li> | ||
<li><a href="https://2010.igem.org/Team:UC_Davis/protocols/gelextraction.html" class="help">Gel extract</a></li></ul></td> | <li><a href="https://2010.igem.org/Team:UC_Davis/protocols/gelextraction.html" class="help">Gel extract</a></li></ul></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td><img src="https://static.igem.org/mediawiki/2010/2/29/RBSplasmidx.jpg"></td> | ||
</tr> | </tr> | ||
</table> | </table> | ||
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<tr> | <tr> | ||
<td><ul><li>Separate fragments through <a href="https://2010.igem.org/Team:UC_Davis/protocols/gelelectrophoresis.html" class="help"> gel electrophoresis</a></li><li><a href="https://2010.igem.org/Team:UC_Davis/protocols/gelextraction.html" class="help">Gel extract</a></li></ul></td> | <td><ul><li>Separate fragments through <a href="https://2010.igem.org/Team:UC_Davis/protocols/gelelectrophoresis.html" class="help"> gel electrophoresis</a></li><li><a href="https://2010.igem.org/Team:UC_Davis/protocols/gelextraction.html" class="help">Gel extract</a></li></ul></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td><img src="https://static.igem.org/mediawiki/2010/c/c5/C0051insert1.jpg"></td> | ||
</tr> | </tr> | ||
</table> | </table> | ||
</td> | </td> | ||
</tr> | </tr> | ||
- | </table><p> | + | </table><p><br /> |
+ | |||
+ | <ul> | ||
+ | <li><a href="https://2010.igem.org/Team:UC_Davis/protocols/ligation.html" class="help">Ligate</a> the RBS and BBa_C0051 fragments together and <a href="https://2010.igem.org/Team:UC_Davis/protocols/transformation.html" class="help">transform</a> on carb plates.</li> | ||
+ | <li>Depending on the outcome of visible colonies on the experimental and control plates, a <a href="https://2010.igem.org/Team:UC_Davis/protocols/pcrscreen.html" class="help">PCR screening</a> may be needed. | ||
+ | </ul><br /> | ||
+ | |||
+ | <img src="https://static.igem.org/mediawiki/2010/b/bb/RBSc0051.jpg"><br /> | ||
+ | |||
+ | Assembly notes | ||
+ | <ul> | ||
+ | <li>It is most practical to cut RBS as a vector because RBS is less than 100 bp long as an insert, making it hard to see and extract on the gel.</li> | ||
+ | <li>The sticky ends from the XbaI and SpeI, when ligated together, will form an 8 bp scar. Thus, XbaI and SpeI will no longer be able to cut here.</li> | ||
+ | </ul><br /> | ||
+ | |||
+ | <a name="stop"><h1>'RBS & BBa_C0051' to Terminator</h1></a> | ||
+ | <ul> | ||
+ | <li><a href="https://2010.igem.org/Team:UC_Davis/protocols/culture.html" class="help">Cultured</a> cells that have parts: 'RBS & BBa_C0051', BBa_B0015</li> | ||
+ | <li><a href="https://2010.igem.org/Team:UC_Davis/protocols/miniprep.html" class="help">Miniprepped</a> 'RBS & BBa_C0051' and BBa_B0015</li> | ||
+ | </ul> | ||
+ | |||
+ | <table border="0" margin="1" width="650px" padding="5px"> | ||
+ | <tr> | ||
+ | <td> | ||
+ | <table border="0" margin="1" width="320px" padding="5px"> | ||
+ | <tr> | ||
+ | <td><img src="https://static.igem.org/mediawiki/2010/9/9a/Stopplasmid.jpg"></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td><ul><li><a href="https://2010.igem.org/Team:UC_Davis/protocols/digestion.html" class="help">Digest</a> with EcoRI & XbaI</li></ul></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td><img src="https://static.igem.org/mediawiki/2010/7/70/Xstopopenvector.jpg"></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td><ul> | ||
+ | <li><a href="https://2010.igem.org/Team:UC_Davis/protocols/gelelectrophoresis.html" class="help">Run on gel</a></li> | ||
+ | <li><a href="https://2010.igem.org/Team:UC_Davis/protocols/gelextraction.html" class="help">Gel extract</a></li></ul></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td><img src="https://static.igem.org/mediawiki/2010/7/70/Xstopopenvector.jpg"></td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </td> | ||
+ | <td> | ||
+ | <table border="0" margin="1" width="320px" padding="5px"> | ||
+ | <tr> | ||
+ | <td><img src="https://static.igem.org/mediawiki/2010/b/bb/RBSc0051.jpg"></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td><ul><li><a href="https://2010.igem.org/Team:UC_Davis/protocols/digestion.html" class="help">Digest</a> with EcoRI & SpeI</li></ul> </td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td><img src="https://static.igem.org/mediawiki/2010/9/94/RBSc0051cut.jpg"></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td><ul><li>Separate fragments through <a href="https://2010.igem.org/Team:UC_Davis/protocols/gelelectrophoresis.html" class="help"> gel electrophoresis</a></li><li><a href="https://2010.igem.org/Team:UC_Davis/protocols/gelextraction.html" class="help">Gel extract</a></li></ul></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td><img src="https://static.igem.org/mediawiki/2010/d/de/RBSc0051insertx.jpg"></td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </td> | ||
+ | </tr> | ||
+ | </table><p><br /> | ||
+ | |||
+ | <ul> | ||
+ | <li><a href="https://2010.igem.org/Team:UC_Davis/protocols/ligation.html" class="help">Ligate</a> the fragments together and <a href="https://2010.igem.org/Team:UC_Davis/protocols/transformation.html" class="help">transform</a> on kan plates.</li> | ||
+ | <li>Depending on the outcome of visible colonies on the experimental and control plates, a <a href="https://2010.igem.org/Team:UC_Davis/protocols/pcrscreen.html" class="help">PCR screening</a> may be needed. | ||
+ | </ul><br /> | ||
+ | |||
+ | <img src="https://static.igem.org/mediawiki/2010/e/eb/RBSc0051stop.jpg"><br /><p> | ||
+ | |||
+ | Assembly notes <br /> | ||
+ | <ul> | ||
+ | <li>It is most practical to cut BBa_B0015 as the vector because it is only 130 bp long. It would be hard to see on a gel. </li> | ||
+ | </ul><br /> | ||
+ | |||
+ | <a name="r0082"><h1>'RBS & BBa_C0051 & stop' to BBa_R0082</h1></a> | ||
+ | <ul> | ||
+ | <li><a href="https://2010.igem.org/Team:UC_Davis/protocols/culture.html" class="help">Cultured</a> cells that have parts: 'RBS & BBa_C0051 & stop', BBa_R0082</li> | ||
+ | <li><a href="https://2010.igem.org/Team:UC_Davis/protocols/miniprep.html" class="help">Miniprepped</a> 'RBS & BBa_C0051& stop' and BBa_R0082</li> | ||
+ | </ul> | ||
+ | |||
+ | <table border="0" margin="1" width="650px" padding="5px"> | ||
+ | <tr> | ||
+ | <td> | ||
+ | <table border="0" margin="1" width="320px" padding="5px"> | ||
+ | <tr> | ||
+ | <td><img src="https://static.igem.org/mediawiki/2010/c/cc/Promoterclosed.jpg"></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td><ul><li><a href="https://2010.igem.org/Team:UC_Davis/protocols/digestion.html" class="help">Digest</a> with SpeI & PstI</li></ul></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td><img src="https://static.igem.org/mediawiki/2010/2/21/Promoterplasmidopenx.jpg"></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td><ul> | ||
+ | <li><a href="https://2010.igem.org/Team:UC_Davis/protocols/gelelectrophoresis.html" class="help">Run on gel</a></li> | ||
+ | <li><a href="https://2010.igem.org/Team:UC_Davis/protocols/gelextraction.html" class="help">Gel extract</a></li></ul></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td><img src="https://static.igem.org/mediawiki/2010/2/21/Promoterplasmidopenx.jpg"></td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </td> | ||
+ | <td> | ||
+ | <table border="0" margin="1" width="320px" padding="5px"> | ||
+ | <tr> | ||
+ | <td><img src="https://static.igem.org/mediawiki/2010/e/eb/RBSc0051stop.jpg"></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td><ul><li><a href="https://2010.igem.org/Team:UC_Davis/protocols/digestion.html" class="help">Digest</a> with XbaI & PstI</li></ul> </td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td><img src="https://static.igem.org/mediawiki/2010/4/46/RBSc0051stopcut.jpg"></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td><ul><li>Separate fragments through <a href="https://2010.igem.org/Team:UC_Davis/protocols/gelelectrophoresis.html" class="help"> gel electrophoresis</a></li><li><a href="https://2010.igem.org/Team:UC_Davis/protocols/gelextraction.html" class="help">Gel extract</a></li></ul></td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td><img src="https://static.igem.org/mediawiki/2010/6/61/RBSc0051stopinsertx.jpg"></td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | </td> | ||
+ | </tr> | ||
+ | </table><p><br /> | ||
+ | |||
+ | <ul> | ||
+ | <li><a href="https://2010.igem.org/Team:UC_Davis/protocols/ligation.html" class="help">Ligate</a> the fragments together and <a href="https://2010.igem.org/Team:UC_Davis/protocols/transformation.html" class="help">transform</a> on carb plates.</li> | ||
+ | <li>Depending on the outcome of visible colonies on the experimental and control plates, a <a href="https://2010.igem.org/Team:UC_Davis/protocols/pcrscreen.html" class="help">PCR screening</a> may be needed. | ||
+ | </ul><br /> | ||
+ | |||
+ | <img src="https://static.igem.org/mediawiki/2010/3/34/Initiatorplasmid.jpg"><br /> | ||
+ | |||
+ | Assembly notes | ||
+ | <ul> | ||
+ | <li>The promoter had to be cut as a vector because it is only 108 bp by itself.</li> | ||
+ | </ul><br /> | ||
</td> | </td> | ||
Line 90: | Line 239: | ||
<ul> | <ul> | ||
<li><a href="#startup" class="help">Starting Up</a></li> | <li><a href="#startup" class="help">Starting Up</a></li> | ||
- | <li><a href="#rbsc0051" class="help"> | + | <li><a href="#rbsc0051" class="help">RBS to c0051</a></li> |
+ | <li><a href="#stop" class="help">'RBS & c0051' to stop</a></li> | ||
+ | <li><a href="#r0082" class="help">'RBS & c0051 & stop' to R0082</a></li> | ||
</ul> </td> | </ul> </td> | ||
</tr> | </tr> |
Latest revision as of 11:35, 19 September 2010
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