TITLE: No Title
AUTHORS: 

[DESCRIPTION]
<p>A protocol to cast a polyacrylamide gel. </p>
<p>Used in for iGEM SDU 2016</p>

[GUIDELINES]
<p><span style="text-decoration: underline;"><strong>20% Separation gel-mix 15 mL</strong></span></p>
<table style="height: 198px;" width="406">
<tbody>
<tr>
<td style="width: 126.515625px;"><span style="font-weight: 400;">10 mL</span></td>
<td style="width: 129.4375px;">
<p><span style="font-weight: 400;">30% Acrylamide/bis</span></p>
</td>
<td style="width: 128.0625px;"> </td>
</tr>
<tr>
<td style="width: 126.515625px;"><span style="font-weight: 400;">3.75 mL</span></td>
<td style="width: 129.4375px;">
<p><span style="font-weight: 400;">1.5M Tris-HCl, pH 8.8</span></p>
</td>
<td style="width: 128.0625px;"> </td>
</tr>
<tr>
<td style="width: 126.515625px;"><span style="font-weight: 400;">150 μL</span></td>
<td style="width: 129.4375px;"><span style="font-weight: 400;">10% SDS</span></td>
<td style="width: 128.0625px;"> </td>
</tr>
<tr>
<td style="width: 126.515625px;"><span style="font-weight: 400;">1.03 mL</span></td>
<td style="width: 129.4375px;"><span style="font-weight: 400;">diH2O</span></td>
<td style="width: 128.0625px;"> </td>
</tr>
<tr>
<td style="width: 126.515625px;"><span style="font-weight: 400;">7.5 μL</span></td>
<td style="width: 129.4375px;"><span style="font-weight: 400;">TEMED</span></td>
<td style="width: 128.0625px;"><span style="font-weight: 400;">ADD JUST BEFORE CASTING!</span></td>
</tr>
<tr>
<td style="width: 126.515625px;"><span style="font-weight: 400;">75 μL</span></td>
<td style="width: 129.4375px;"><span style="font-weight: 400;">10% APS</span></td>
<td style="width: 128.0625px;"><span style="font-weight: 400;">ADD JUST BEFORE CASTING</span></td>
</tr>
</tbody>
</table>
<p> </p>
<p> </p>
<p> </p>
<p><strong>4% Stacking gel-mix 15 mL</strong></p>
<table style="height: 198px;" width="406">
<tbody>
<tr>
<td style="width: 126.515625px;"><span style="font-weight: 400;">1.98 mL</span></td>
<td style="width: 129.4375px;">
<p><span style="font-weight: 400;">30% Acrylamide/bis</span></p>
</td>
<td style="width: 128.0625px;"> </td>
</tr>
<tr>
<td style="width: 126.515625px;"><span style="font-weight: 400;">3.78 mL</span></td>
<td style="width: 129.4375px;">
<p><span style="font-weight: 400;">0.5M Tris-HCl, pH6.8</span></p>
</td>
<td style="width: 128.0625px;"> </td>
</tr>
<tr>
<td style="width: 126.515625px;"><span style="font-weight: 400;">150 μL</span></td>
<td style="width: 129.4375px;"><span style="font-weight: 400;">10% SDS</span></td>
<td style="width: 128.0625px;"> </td>
</tr>
<tr>
<td style="width: 126.515625px;"><span style="font-weight: 400;">9 mL</span></td>
<td style="width: 129.4375px;"><span style="font-weight: 400;">diH2O</span></td>
<td style="width: 128.0625px;"> </td>
</tr>
<tr>
<td style="width: 126.515625px;"><span style="font-weight: 400;">15 μL</span></td>
<td style="width: 129.4375px;"><span style="font-weight: 400;">TEMED</span></td>
<td style="width: 128.0625px;"><span style="font-weight: 400;">ADD JUST BEFORE CASTING!</span></td>
</tr>
<tr>
<td style="width: 126.515625px;"><span style="font-weight: 400;">75 μL</span></td>
<td style="width: 129.4375px;"><span style="font-weight: 400;">10% APS</span></td>
<td style="width: 128.0625px;"><span style="font-weight: 400;">ADD JUST BEFORE CASTING</span></td>
</tr>
</tbody>
</table>
<p> </p>
<p><strong>X% Separation gel-mix 15 mL</strong></p>
<table style="height: 198px;" width="406">
<tbody>
<tr>
<td style="width: 126.515625px;"><span style="font-weight: 400;">0.5 * X mL</span></td>
<td style="width: 129.4375px;">
<p><span style="font-weight: 400;">30% Acrylamide/bis</span></p>
</td>
<td style="width: 128.0625px;"> </td>
</tr>
<tr>
<td style="width: 126.515625px;"><span style="font-weight: 400;">3.75 mL</span></td>
<td style="width: 129.4375px;">
<p><span style="font-weight: 400;">1.5M Tris-HCl, pH 8.8</span></p>
</td>
<td style="width: 128.0625px;"> </td>
</tr>
<tr>
<td style="width: 126.515625px;"><span style="font-weight: 400;">150 μL</span></td>
<td style="width: 129.4375px;"><span style="font-weight: 400;">10% SDS</span></td>
<td style="width: 128.0625px;"> </td>
</tr>
<tr>
<td style="width: 126.515625px;"><span style="font-weight: 400;">11.03-(0.5*X) mL</span></td>
<td style="width: 129.4375px;"><span style="font-weight: 400;">diH2O</span></td>
<td style="width: 128.0625px;"> </td>
</tr>
<tr>
<td style="width: 126.515625px;"><span style="font-weight: 400;">7.5 μL</span></td>
<td style="width: 129.4375px;"><span style="font-weight: 400;">TEMED</span></td>
<td style="width: 128.0625px;"><span style="font-weight: 400;">ADD JUST BEFORE CASTING!</span></td>
</tr>
<tr>
<td style="width: 126.515625px;"><span style="font-weight: 400;">75 μL</span></td>
<td style="width: 129.4375px;"><span style="font-weight: 400;">10% APS</span></td>
<td style="width: 128.0625px;"><span style="font-weight: 400;">ADD JUST BEFORE CASTING</span></td>
</tr>
</tbody>
</table>
<p> </p>
<p><strong>2x SDS-PAGE Buffer</strong></p>
<table style="height: 198px;" width="406">
<tbody>
<tr>
<td style="width: 126.515625px;"><span style="font-weight: 400;">3.75 mL</span></td>
<td style="width: 129.4375px;">
<p><span style="font-weight: 400;">Tris-HCl pH 6.8, 0.5M</span></p>
</td>
<td style="width: 128.0625px;"> </td>
</tr>
<tr>
<td style="width: 126.515625px;"><span style="font-weight: 400;">24.0 mL</span></td>
<td style="width: 129.4375px;">
<p><span style="font-weight: 400;">Glycerol</span></p>
</td>
<td style="width: 128.0625px;"> </td>
</tr>
<tr>
<td style="width: 126.515625px;"><span style="font-weight: 400;">300 μL</span></td>
<td style="width: 129.4375px;"><span style="font-weight: 400;">1.0% Bromophenol blue eller xylene cyanol (MEGET Lidt )</span></td>
<td style="width: 128.0625px;"> </td>
</tr>
<tr>
<td style="width: 126.515625px;"><span style="font-weight: 400;">6.0 mL</span></td>
<td style="width: 129.4375px;"><span style="font-weight: 400;">10% SDS</span></td>
<td style="width: 128.0625px;"> </td>
</tr>
<tr>
<td style="width: 126.515625px;"><span style="font-weight: 400;">-&gt; 30 mL</span></td>
<td style="width: 129.4375px;"><span style="font-weight: 400;">diH</span><sub><span style="font-weight: 400;">2</span></sub><span style="font-weight: 400;">O</span></td>
<td style="width: 128.0625px;"> </td>
</tr>
</tbody>
</table>
<p> </p>
<p> </p>
<p><strong>Acrylamide/Bis 30%</strong></p>
<table style="height: 198px;" width="406">
<tbody>
<tr>
<td style="width: 126.515625px;"><span style="font-weight: 400;">87.60 g</span></td>
<td style="width: 129.4375px;">
<p><span style="font-weight: 400;">Acrylamide (29.2g / 100mL )</span></p>
</td>
<td style="width: 128.0625px;"> </td>
</tr>
<tr>
<td style="width: 126.515625px;"><span style="font-weight: 400;">2.40 g</span></td>
<td style="width: 129.4375px;">
<p><span style="font-weight: 400;">N'N'-bis-methylene-acrylamide</span></p>
</td>
<td style="width: 128.0625px;"> </td>
</tr>
<tr>
<td style="width: 126.515625px;"><span style="font-weight: 400;">-&gt; 300 mL</span></td>
<td style="width: 129.4375px;"><span style="font-weight: 400;">diH</span><sub><span style="font-weight: 400;">2</span></sub><span style="font-weight: 400;">O</span></td>
<td style="width: 128.0625px;"> </td>
</tr>
<tr>
<td style="width: 126.515625px;"><span style="font-weight: 400;">Filter and store at 4ºC</span></td>
<td style="width: 129.4375px;"> </td>
<td style="width: 128.0625px;"> </td>
</tr>
<tr>
<td style="width: 126.515625px;"> </td>
<td style="width: 129.4375px;"> </td>
<td style="width: 128.0625px;"> </td>
</tr>
</tbody>
</table>
<p> </p>
<p> </p>
<p><strong>1.5 M Tris-HCl, pH 8.8 (150 ml)</strong></p>
<table style="height: 198px;" width="406">
<tbody>
<tr>
<td style="width: 126.515625px;"><span style="font-weight: 400;">27.23 g</span></td>
<td style="width: 129.4375px;">
<p><span style="font-weight: 400;">Tris-base (18.15 g/100 ml)</span></p>
</td>
<td style="width: 128.0625px;"> </td>
</tr>
<tr>
<td style="width: 126.515625px;"><span style="font-weight: 400;">80 mL</span></td>
<td style="width: 129.4375px;">
<p><span style="font-weight: 400;">diH</span><sub><span style="font-weight: 400;">2</span></sub><span style="font-weight: 400;">O</span></p>
</td>
<td style="width: 128.0625px;"> </td>
</tr>
<tr>
<td style="width: 126.515625px;">
<p><span style="font-weight: 400;">Adjust to pH 8.8 with 6N HCl</span></p>
</td>
<td style="width: 129.4375px;"> </td>
<td style="width: 128.0625px;"> </td>
</tr>
<tr>
<td style="width: 126.515625px;">-&gt; 150 mL</td>
<td style="width: 129.4375px;">diH<sub>2</sub>O</td>
<td style="width: 128.0625px;"> </td>
</tr>
<tr>
<td style="width: 126.515625px;">
<p>store at 4ºC</p>
</td>
<td style="width: 129.4375px;"> </td>
<td style="width: 128.0625px;"> </td>
</tr>
</tbody>
</table>
<p> </p>
<p> </p>
<p><strong>0.5 M Tris-HCl, pH 6.8</strong></p>
<table style="height: 198px;" width="406">
<tbody>
<tr>
<td style="width: 126.515625px;"><span style="font-weight: 400;">6 g</span></td>
<td style="width: 129.4375px;">
<p><span style="font-weight: 400;">Tris-base (18.15 g/100 ml)</span></p>
</td>
<td style="width: 128.0625px;"> </td>
</tr>
<tr>
<td style="width: 126.515625px;"><span style="font-weight: 400;">60 mL</span></td>
<td style="width: 129.4375px;">
<p><span style="font-weight: 400;">diH</span><sub><span style="font-weight: 400;">2</span></sub><span style="font-weight: 400;">O</span></p>
</td>
<td style="width: 128.0625px;"> </td>
</tr>
<tr>
<td style="width: 126.515625px;">
<p><span style="font-weight: 400;">Adjust to pH 6.87 with 6N HCl</span></p>
</td>
<td style="width: 129.4375px;"> </td>
<td style="width: 128.0625px;"> </td>
</tr>
<tr>
<td style="width: 126.515625px;">-&gt; 100 mL</td>
<td style="width: 129.4375px;">diH<sub>2</sub>O</td>
<td style="width: 128.0625px;"> </td>
</tr>
<tr>
<td style="width: 126.515625px;">
<p>store at 4ºC</p>
</td>
<td style="width: 129.4375px;"> </td>
<td style="width: 128.0625px;"> </td>
</tr>
</tbody>
</table>
<p> </p>
<p> </p>
<p><strong>10% APS</strong></p>
<table style="height: 102px;" width="401">
<tbody>
<tr style="height: 42px;">
<td style="width: 124.796875px; height: 42px;"><span style="font-weight: 400;">0.10 g</span></td>
<td style="width: 128.328125px; height: 42px;">
<p><span style="font-weight: 400;">Ammonium persulfate</span></p>
</td>
<td style="width: 125.90625px; height: 42px;"> </td>
</tr>
<tr style="height: 42px;">
<td style="width: 124.796875px; height: 42px;"><span style="font-weight: 400;">1 mL</span></td>
<td style="width: 128.328125px; height: 42px;">
<p><span style="font-weight: 400;">diH</span><sub><span style="font-weight: 400;">2</span></sub><span style="font-weight: 400;">O</span></p>
</td>
<td style="width: 125.90625px; height: 42px;"> </td>
</tr>
</tbody>
</table>
<p> </p>
<p><strong>10% SDS</strong></p>
<table style="height: 198px;" width="406">
<tbody>
<tr>
<td style="width: 126.515625px;"><span style="font-weight: 400;">10.00 g</span></td>
<td style="width: 129.4375px;">
<p><span style="font-weight: 400;">SDS</span></p>
</td>
<td style="width: 128.0625px;"> </td>
</tr>
<tr>
<td style="width: 126.515625px;"><span style="font-weight: 400;">60 mL</span></td>
<td style="width: 129.4375px;">
<p><span style="font-weight: 400;">diH</span><sub><span style="font-weight: 400;">2</span></sub><span style="font-weight: 400;">O</span></p>
</td>
<td style="width: 128.0625px;"> </td>
</tr>
<tr>
<td style="width: 126.515625px;">
<p><span style="font-weight: 400;">Dissolve with gentle stirring</span></p>
</td>
<td style="width: 129.4375px;"> </td>
<td style="width: 128.0625px;"> </td>
</tr>
<tr>
<td style="width: 126.515625px;">-&gt; 100 mL</td>
<td style="width: 129.4375px;">diH<sub>2</sub>O</td>
<td style="width: 128.0625px;"> </td>
</tr>
<tr>
<td style="width: 126.515625px;">
<p> </p>
</td>
<td style="width: 129.4375px;"> </td>
<td style="width: 128.0625px;"> </td>
</tr>
</tbody>
</table>
<p> </p>

[STEPS]
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