Team:St Andrews/Procedure

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        <header class="jumbotron subhead" id="overview">
 
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            <h1>Lab Book</p>
 
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<p></p>
 
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<h2>Procedure</h2>
 
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            </header>
 
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    <li><a href="#tab1" data-toggle="tab">ω-3 Synthesis</a></li>
 
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    <li><a href="#tab2" data-toggle="tab">Metal-binding peptides</a></li>
 
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<BR>&nbsp;</BR>
 
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<p><span class="label label-info"><a href="#Primers2"><font color="white">Primers</font></a></span>&nbsp;<span class="label label-info"><a href="#Digestion2"><font color="white">Digestion</font></a></span>&nbsp;<span class="label label-info"><a href="#Transformation2"><font color="white">Transformation</font></a></span></p>
 
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<BR>&nbsp;</BR>
 
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<a name="Primers2"><span class="label label-info">Primers</span></a>
 
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<BR>&nbsp;</BR>
 
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<p>All primers are notated 5' to 3'.</p>
 
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<p>Please refer to the <a href="https://2012.igem.org/Team:St_Andrews/Lab-book#Primer annealing"><strong>Primer annealing lab book entry</strong></a> for further detail.</p>
 
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<p> </p>
 
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  <a class="btn dropdown-toggle" data-toggle="dropdown" href="#">
 
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  Ni forward
 
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    <div class="span3">AATTCCATCATCACCATCACCACC</ul>
 
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    Ni reverse
 
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    <span class="caret"></span>
 
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  <p><div class="span3">TCGAGGTGGTGATCGTGATGATGG</div></p> 
 
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    Ni<sub>2</sub> forward
 
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      <div class="span4">AATTCATGTGTACAACATGCGGTTGCGGTGAAGGCC</div>
 
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    Ni<sub>2</sub> reverse
 
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    <span class="caret"></span>
 
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  </a>
 
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      <div class="span4">TCGAGGCCTTCACCGCAACCGCATGTTGTACACATG</div>
 
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    Pd forward
 
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    <span class="caret"></span>
 
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      <div class="span3">AGCGTGACCCAGAACAAATAT</div>
 
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  <a class="btn dropdown-toggle" data-toggle="dropdown" href="#">
 
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    Pd reverse
 
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      <div class="span3">ATATTTGTTCTGGGTCACGCT</div>
 
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  <a class="btn dropdown-toggle" data-toggle="dropdown" href="#">
 
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    Pt forward
 
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<div class="span3">GATCGCACCAGCACCTGGCGC</div>
 
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    Pt reverse
 
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  <div class="span3">GCGCCAGGTGCTGGTGCGATC</div>
 
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</div></p>
 
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<BR>&nbsp;</BR>
 
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<p>For primer annealing in the PCR, the primer sequences were combined in the following way:</p>
 
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<p>Please refer to the <a href="https://2012.igem.org/Team:St_Andrews/Lab-book"><strong>Primer annealing lab book entry</strong></a> for further detail.</p>
 
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<li><strong>GST Forward</strong> and <strong>Ni Reverse</strong></li>
 
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<li><strong>GST Forward</strong> and <strong>Ni<sub>2</sub> Reverse</strong></li>
 
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<li><strong>GST Forward</strong> and <strong>Pd Reverse</strong></li>
 
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<li><strong>GST Forward</strong> and <strong>Pt Reverse</strong></li>
 
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<BR>&nbsp;</BR>
 
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<a name="Digestion2"><span class="label label-info">Digestion</span></a>
 
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<BR>&nbsp;</BR>
 
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<p>pGEX-6P-1 was cut using EcoR1(954) and Xho1 (975)</p>
 
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<BR>&nbsp;</BR>
 
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<a name="Transformation2"><span class="label label-info">Transformation</span></a>
 
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<p>pGEX vector with insert transformed into DH5α E. coli cells to replicate.</p>
 
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<p>pGEX vector with G-Block insert transformed into DH5α E. coli cells to replicate.</p>
 
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<BR>&nbsp;</BR>
 
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<p>pGEX vector with insert transformed into BL21 E. coli cells for protein expression.</p>
 
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<p>pGEX vector with G-Block insert transformed into BL21 E. coli cells for protein expression.</p>
 
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Latest revision as of 11:51, 23 September 2012