Team:UC Chile2/General Protocols

From 2012.igem.org

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{{UC_Chile4}}
{{UC_Chile4}}
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<div id="Media">
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<div id=" Growth Media">
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== Growth Media ==
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</div>
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== Media ==
 
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</div>
 
<h1>LB media</h1>
<h1>LB media</h1>
<ul>
<ul>
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<li>Adjust pH to 7.5 with NaOH 1M</li>
<li>Adjust pH to 7.5 with NaOH 1M</li>
</ul>
</ul>
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<div id="Buffers">
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== Buffers ==
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</div>
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<h1>5X Gibson Isothermal Buffer</h1>
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<ul>
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<li>25% PEG MW 8000</li>
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<li>500mM Tris HCl pH 7.5</li>
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<li>50mM CaCl</li>
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<li>50mM DTT</li>
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<li>1mM dATP</li>
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<li>1mM dTTP</li>
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<li>1mM dGTP</li>
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<li>1mM dCTP</li>
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<li>5mM NAD+</li>
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<li>H20 nuclease free</li>
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<p>For a 2 mL stock (equivalent to 20 100uL 5X isothermal buffers)</p>
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<li>0.5g PEG MW 8000</li>
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<li>1.5mL Tris-HCl pH 7.5 1M</li>
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<p>Swivel for 30 minutes in a orbital oscillator </p>
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<li>50uL CaCl 2M</li>
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<li>100uL DTT 1M</li>
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<li>20uL dATP 100mM</li>
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<li>20uL dTTP 100mM</li>
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<li>20uL dGTP 100mM</li>
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<li>20uL dCTP 100mM</li>
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<li>200uL NAD+ 200mM</li>
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<li>H20 nuclease free up to 2mL</li>
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<p>Alicuot into 100uL stocks</p>
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<b>Store at -80°C</b>
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</ul>
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<div id="DNA assembly">
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== DNA assembly ==
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</div>
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<h1>Gibson Assembly</h1>

Revision as of 02:40, 12 September 2012

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