Team:Calgary/Notebook/Protocols/GelExtraction

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<div id="Banner"><h1>Gel Extraction</h1></div>
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<h1>Gel Extraction</h1>

<p class="noIndent">This protocol is utilized in accordance to the manufacturer's protocol from Omega E.Z.N.A (EaZy Nucleic Acid Isolation)</p>
<h2>Reagents and Materials</h2>
<ul>
<li>Omega E.Z.N.A (EaZy Nucleic Acid Isolation) kit</li>
<li>Agarose gel</li>
<li>Blade</li>
</ul>
<h2>Protocol</h2>
<ol>
<li>Place gel on the UV box</li>
<li>Carefully extract the fragment suspended in the gel</li>
<li>Mass gel fragments</li>
<li>Place fragment into a 1.5 mL tube and add 4 µL of H<sub>2</sub>O</li>
<li>Volume of water added to volume of gel is 200% however if fragment it small 1 mL of water will suffice</li>
<li>Remove H<sub>2</sub>O</li>
<li>Add equal amounts of H2O and Binding Buffer (XP2) to the gel</li>
<li>Incubate mixture at 55°C for 7 mins</li>
<li>Mix with vortex for 2 mins</li>
<li>Place in the HiBind DNA Mini Column in the 2 mL tube</li>
<li>Add 700 µL at 10,000xg for 1 min</li>
<li>Discard liquid</li>
<li>Add 300 µL Binding Buffer (XP2) into the HiBind DNA Mini Column and spin down at 10,000xg for 1 min</li>
<li>Discard liquid</li>
<li>Wash the column with 700 µL of SPW buffer with added ethanol and spin down at 10,000xg for 1 min</li>
<li>Discard liquid</li>
<li>Wash the column with 700 µL of SPW buffer again and spin down at 10,000xg for 1 min</li>
<li>Discard the liquid</li>
<li>Spin down the column at 13,000xg for 1 min to dry the column</li>
<li>Elute in 50 µL of H<sub>2</sub>O and wait 1 min</li>
<li>Spin down the column at 13,000xg for 1 min to dry the column</li>
<li>Use a spectrophotometer to measure the concentration and the purity of your plasmid</li>
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