Team:UC Davis/notebook/initiator.html

From 2010.igem.org

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<img src="https://static.igem.org/mediawiki/2010/e/eb/RBSc0051stop.jpg"><br /><p>
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<a name="r0082"><h1>'RBS & BBa_C0051 & stop' to BBa_R0082</h1></a>
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<ul>
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<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>
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<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>
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<table border="0" margin="1" width="650px" padding="5px">
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      <td><img src="https://static.igem.org/mediawiki/2010/c/cc/Promoterclosed.jpg"></td>
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      <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>
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      <td><img src="https://static.igem.org/mediawiki/2010/d/d2/Promoterplasmidopen.jpg"></td>
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        <li><a href="https://2010.igem.org/Team:UC_Davis/protocols/gelelectrophoresis.html" class="help">Run on gel</a></li>
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        <li><a href="https://2010.igem.org/Team:UC_Davis/protocols/gelextraction.html" class="help">Gel extract</a></li></ul></td>
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      <td><img src="https://static.igem.org/mediawiki/2010/d/d2/Promoterplasmidopen.jpg"></td>
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  <table border="0" margin="1" width="320px" padding="5px">
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      <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>
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      <td><img src="https://static.igem.org/mediawiki/2010/4/46/RBSc0051stopcut.jpg"></td>
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      <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>
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      <td><img src="https://static.igem.org/mediawiki/2010/6/61/RBSc0051stopinsertx.jpg"></td>
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</table><p><br />
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<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>
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<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.
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<img src="https://static.igem.org/mediawiki/2010/3/34/Initiatorplasmid.jpg"><br />
                            
                            
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Revision as of 10:51, 19 September 2010

The Initiator

Starting Up

  • Hydrated parts: BBa_B0034, BBa_C0051, BBa_B0015, BBa_R0082.
  • Transformed these parts into DH5α competent cells.


RBS to BBa_C0051


  • Ligate the RBS and BBa_C0051 fragments together and transform on carb plates.
  • Depending on the outcome of visible colonies on the experimental and control plates, a PCR screening may be needed.


Assembly notes
  • 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.
  • 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.

'RBS & BBa_C0051' to Terminator
  • Cultured cells that have parts: 'RBS & BBa_C0051', BBa_B0015
  • Miniprepped 'RBS & BBa_C0051' and BBa_B0015


  • Ligate the fragments together and transform on kan plates.
  • Depending on the outcome of visible colonies on the experimental and control plates, a PCR screening may be needed.


'RBS & BBa_C0051 & stop' to BBa_R0082

  • Cultured cells that have parts: 'RBS & BBa_C0051 & stop', BBa_R0082
  • Miniprepped 'RBS & BBa_C0051& stop' and BBa_R0082


  • Ligate the fragments together and transform on carb plates.
  • Depending on the outcome of visible colonies on the experimental and control plates, a PCR screening may be needed.


We would like to take a moment to thank all of our sponsors for their very generous donations, as we could not have done this without your help!

We would also like to thank and acknowledge:
Our Advisors
Marc Facciotti
Ilias Tagkopoulos
Technical Guidance
David Larsen
Andrew Yao
Visiting iGEMer
Jia Li of Zhejiang University (TEAM ZJU-China)
cI Promoter Screen
Drew Endy - Stanford
Thomas Schneider - NIH
Want to sponsor us? Send an email to mtfacciotti@ucdavis.edu to discuss various ways you can help! :)

Retrieved from "http://2010.igem.org/Team:UC_Davis/notebook/initiator.html"