Team:UC Chile2/General Protocols

From 2012.igem.org

(Difference between revisions)
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<h1>LB media</h1>
<h1>LB media</h1>
<ul>
<ul>
-
<p>Final Concentrations</p>
+
<p>Final concentrations:</p>
<li>1% (w/v) tryptone</li>
<li>1% (w/v) tryptone</li>
<li>0.5% (w/v) yeast extract</li>
<li>0.5% (w/v) yeast extract</li>
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<h1>SOB media</h1>
<h1>SOB media</h1>
<ul>
<ul>
-
<p>Final Concentrations</p>
+
<p>Final concentrations:</p>
<li>0.5% (w/v) yeast extract</li>
<li>0.5% (w/v) yeast extract</li>
<li>2% (w/v) tryptone</li>
<li>2% (w/v) tryptone</li>
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== Buffers ==
== Buffers ==
</div>
</div>
-
<h1>5X Gibson Isothermal Buffer</h1>
+
<h1>5X Gibson Assembly Isothermal Buffer</h1>
<ul>
<ul>
-
<p>Final Concentrations</p>
+
<p>Final Concentrations:</p>
<li>25% PEG MW 8000</li>
<li>25% PEG MW 8000</li>
<li>500mM Tris HCl pH 7.5</li>
<li>500mM Tris HCl pH 7.5</li>
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<p>Alicuot into 100uL stocks</p>
<p>Alicuot into 100uL stocks</p>
<b>Store at -80°C</b>
<b>Store at -80°C</b>
 +
</ul>
 +
<h1>1.33X Gibson Assembly Master Mix</h1>
 +
<ul>
 +
<p>For a 375ul 1.33X Gibson Assembly Master Mix</p>
 +
<li>100uL 5X Gibson Assembly Isothermal Buffer</li>
 +
<li>6.25uL Phusion Polymerase XXX U/uL (cat N° F-350S) from Thermo Scientific </li>
 +
<li>2uL Epicentre T5 Exonuclease 1U/ul(cat N° T5E4111K) from Illumina</li>
 +
<p>It is important to dilute the T5 Exonuclease stock from 10U/uL to 1U/ul to measure the accurate volume correctly</p>
 +
<li>50uL Taq DNA Ligase 2000 U/uL (cat N° M208S) from NEB</li>
</ul>
</ul>
<div id="DNA assembly">
<div id="DNA assembly">

Revision as of 02:54, 12 September 2012

Project: Luxilla - Pontificia Universidad Católica de Chile, iGEM 2012