Team:SDU-Denmark/labwork/Protocols/3A

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</br>
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<table>
<tr>
	
<td><a href="https://2012.igem.org/Team:SDU-Denmark/labwork/Protocols">mRNA Isolation</a></td>
<td><a href="https://2012.igem.org/Team:SDU-Denmark/labwork/Protocols/PCR">PCR</a></td>
<td><a href="https://2012.igem.org/Team:SDU-Denmark/labwork/Protocols/Miniprep">Miniprep</a></td>
<td><a href="https://2012.igem.org/Team:SDU-Denmark/labwork/Protocols/Checkdigest">Check Digest</a></td>
</tr>
<tr>
<td><a href="https://2012.igem.org/Team:SDU-Denmark/labwork/Protocols/3A"><b>3A-Assembly</b></a></td>
<td>Content</td>
<td>Content</td>
<td>Content</td>
</tr>
<tr>
<td>Content</td>
<td>Content</td>
<td>Content</td>
<td>Content</td>
</tr>
</table>


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<h2> 3A-Assembly -<i>Ligation of 2 parts into a vector</i> </h2>


<p>
<ol>
<li>PCR step; see PCR protocol</li>
<li>Digestion</li>
<i>BSA is only needed for conventional restriction enzymes, NOT fast digest</i>
<ol type="i">
<li>Digest step for Plasmid Backbone</li>
<ul type="square">
<b>if sample comes from miniprep</b>
<li>Enzyme Master Mix for Plasmid Backbone (25ul tota</li>
<ul>
<li>5 ul Fastdigest greenbuffer / NEB Buffer 2</li>
<li>0.5 ul BSA</li>
<li>0.5 ul EcoRI-H</li>
<li>0.5 ul PstI</li>
<li>19 ul dH20 (0,5 ul less when using BSA)</li>

</ul>
</ul>
</ol>  

<li></li>
<li></li>
<li></li>
<li></li>
<li></li>

</p>


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