Difference between revisions of "Team:Wageningen UR/Parts"

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{{Wageningen_UR/menu}}
 
{{Wageningen_UR/menu}}
 
<html>
 
<html>
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<table>
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  <tr>
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    <th>BioBrick
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name</th>
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    <th>Basic/Composite/Device</th>     
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    <th>Designer</th>
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  </tr>
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  <tr>
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    <td>BBa_K1913000</td>
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    <td>Basic</td>
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    <td>Lisa</td>
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  </tr>
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</table>
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<p>
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When we started making the constructs for the <a “href=https://2016.igem.org/Team:Wageningen_UR/Description/Biocontainment#SAA">Cas9 kill switch</a>, two approaches were taken: one was taking the Cas9 that is available in the iGEM registry (BBa_K1218011) as a starting point for making mutations and expressing Cas9. The other approach was starting with pdCas9 (Addgene plasmid # 46569). While cloning, it proved to be difficult to transfer BBa_K1218011 to another backbone (see also the <a “href=https://2016.igem.org/Team:Wageningen_UR/Notebook/Cas9">notebook</a>). Furthermore, when cultures transformed with BBa_K1218011 were checked for Cas9 expression with SDS-PAGE, no convincing Cas9 band could be observed. Because of time limitations we decided to continue working with the Addgene construct for making the mutations, and chose an established system for protein expression. For that reason, no Cas9-biobricks were submitted. Furthermore, the pEVOL construct containing an aminoacyl-synthetase and a tRNA for introducing BipA in response to the TAG stopcodon were isolated from a strain kindly received from George Church (described in <a “href=http://www.nature.com/nature/journal/v518/n7537/full/nature14121.html">Mandel <i>et al.</i>, 2015</a>). The MTA that was signed to receive the strain does not allow for redistribution. 
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</p>
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<section id="biobrick1">
 
<section id="biobrick1">
 
<h1><b>BioBrick 1</b></h1>
 
<h1><b>BioBrick 1</b></h1>

Revision as of 15:28, 15 October 2016

Wageningen UR iGEM 2016

 

BioBrick name Basic/Composite/Device Designer
BBa_K1913000 Basic Lisa

When we started making the constructs for the Cas9 kill switch, two approaches were taken: one was taking the Cas9 that is available in the iGEM registry (BBa_K1218011) as a starting point for making mutations and expressing Cas9. The other approach was starting with pdCas9 (Addgene plasmid # 46569). While cloning, it proved to be difficult to transfer BBa_K1218011 to another backbone (see also the notebook). Furthermore, when cultures transformed with BBa_K1218011 were checked for Cas9 expression with SDS-PAGE, no convincing Cas9 band could be observed. Because of time limitations we decided to continue working with the Addgene construct for making the mutations, and chose an established system for protein expression. For that reason, no Cas9-biobricks were submitted. Furthermore, the pEVOL construct containing an aminoacyl-synthetase and a tRNA for introducing BipA in response to the TAG stopcodon were isolated from a strain kindly received from George Church (described in Mandel et al., 2015). The MTA that was signed to receive the strain does not allow for redistribution.

BioBrick 1

YOUR TEXT HERE
Link to favourite biobrick should link to Wageningen_UR/Basic_Part
Link to set of favourite biobricks should link to Wageningen_UR/Part_Collection

BioBrick 2

BioBrick 3