Team:Cambridge-JIC/Constructs/35S-HBRmRFP

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<div align="center"><a href="https://2014.igem.org/wiki/index.php?title=Team:Cambridge-JIC/Constructs/35S-HBRmRFP&action=edit">Edit this page.</a></div>
<div align="center"><a href="https://2014.igem.org/wiki/index.php?title=Team:Cambridge-JIC/Constructs/35S-HBRmRFP&action=edit">Edit this page.</a></div>
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<h1> Gal4 - Hap1GR - H2BmRFP</h1>
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<h1> 35S-H2BmRFP</h1>
<h2> Design </h2>
<h2> Design </h2>
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<h4> Aim of the construct </h4>
<h4> Aim of the construct </h4>
<ul>  
<ul>  
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<li> We'll try out 35s->GAL4->HAP1GR->H2BmRFP. </li>
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<li> This is a simple construct for me (Hugh) to check my practical skills. Apparently it has worked for others in the past.</li>
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<li> Since H2BmRFP has not been tried in Mp, we'll also try 35s->H2BmRFP. A positive control for lab technique will be 35s->Venus N7</li>
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<li> We'll also use FF's original plasmid to try 35s-Hap1GR-mRFP, as a control for the Hap1GR. I expect low transformation rate as in the original plasmid, we're using 35s as promoter for HygR selection marker.
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<li> Not sure whether 35s->Gal4 has been tried (check with BP), so 35s-Gal4-venus n7 is doable with BP's construct as a control </li>
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<li> As another control, we'll try 35s-Gal4-H2mRFP </li>
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</ul>
</ul>
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</div>
</div>
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<h4> End goal plasmid </h4>
<h4> End goal plasmid </h4>
<ul>  
<ul>  
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<li> <img src="https://static.igem.org/mediawiki/2014/f/fd/Cambridge-JIC_constructs_GAL4-HAP1GR-H2mRFP.png" width = "100%"> </li>
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<li> <!--<img>--> </li>
</ul>
</ul>
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</div>
</div>
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<h2> Experimentation (up to date)</h2>
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<h2> Experiments (to date)</h2>
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<div id="PCR" align="left">
<div id="PCR" align="left">
<h4>PCR</h4>
<h4>PCR</h4>
Attempt 1:
Attempt 1:
<ul>
<ul>
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<li>Start date/time: 24/07, 11:08</li>
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<li>Start date/time: 30/07, morning</li>
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<li>End date/time:24/07, 1550</li>
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<li>End date/time:30/07, morning</li>
<li> UIDs of primers/plasmids used:  
<li> UIDs of primers/plasmids used:  
<ul>
<ul>
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<li> A1, P33, P34</li>
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<li> A1(DEXX3), P42(35S-H2BF), P41(nosT-mRFP-B)</li>
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<li> A1, P35, P36</li>
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<li> B14(35S-rLTI), P17(P1_B), P15(nosT-F)</li>
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<li> A2, P37, P38</li>
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<li> B14(35S-rLTI), P16(P1_F), P14(35S-B)</li>
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<li> A2, P39, P40 </ul></li>
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</ul></li>
<li><a href="https://docs.google.com/spreadsheets/d/1RwA7YHMnZ4a0M67RMbegWU5ee3MJRo62qOcBJj537yg/edit#gid=1660015451">PCR Spreadsheet (on google drive)</a></li>
<li><a href="https://docs.google.com/spreadsheets/d/1RwA7YHMnZ4a0M67RMbegWU5ee3MJRo62qOcBJj537yg/edit#gid=1660015451">PCR Spreadsheet (on google drive)</a></li>
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<li><a href="http://google.com/GEL_27_07_2014">Gel 27/07/2014</a></li>
 
</ul>
</ul>
</div>
</div>
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</ul>
</ul>
</div>
</div>
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<div id="Spore preparation" align="left">
<div id="Spore preparation" align="left">
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<h4>Spore preparation</h4>
<h4>Spore preparation</h4>
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Attempt 1:
Attempt 1:
<ul>
<ul>
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<div id="Spore transformation" align="left">
<div id="Spore transformation" align="left">
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<h4> Spore transformation</h4>
<h4> Spore transformation</h4>
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Attempt 1:
Attempt 1:
<ul>
<ul>
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<div id="Evaluation" align="left">
<div id="Evaluation" align="left">
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<h2> Evaluation</h2>
<h2> Evaluation</h2>
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<ul>
<ul>
<li>Start date/time:</li>
<li>Start date/time:</li>

Latest revision as of 13:53, 31 July 2014