Team:Calgary/Sandbox/Notebook/Protocols/AgaroseGelElectrophoresis

From 2013.igem.org

(Difference between revisions)
(Created page with "{{Team:Calgary/Main}} <html> <section id="Content"> <h1>Agarose Gel Electrophoresis</h1> <p><b>Reagents and Materials</b></p> <p>1X TA </p> <p>Graduated Cylinder</p> ...")
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<h1>Agarose Gel Electrophoresis</h1>
<h1>Agarose Gel Electrophoresis</h1>
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<p><b>Reagents and Materials</b></p>
+
TITLE: Agarose Gel Electrophoresis
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   <p>1X TA </p>
+
 
-
     <p>Graduated Cylinder</p>
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<h2>Reagents and Materials<h2>
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     <p>125 mL flask </p>
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<ul>
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     <p>Agarose </p>
+
   <li>1X TA </li>
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     <p>Gel Pouring Tray </p>
+
     <li>Graduated Cylinder</li>
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     <p>Tape </p>
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     <li>125 mL flask </li>
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     <p>Gel rig </p>
+
     <li>Agarose </li>
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     <p>Red Safe </p>
+
     <li>Gel Pouring Tray </li>
-
  <p><b>Protocol</b></p>
+
     <li>Tape </li>
-
     <p>Measure out 100mL of 1x TAE buffer;</p>
+
     <li>Gel rig </li>
-
     <p>Transfer buffer to 125 mL flask;</p>
+
     <li>Red Safe </li>
-
     <p>Weigh out enough agarose to make a 1% gel (in our case 1.0 g of agarose was the right amount);</p>
+
</ul>
-
     <p> Transfer agarose to 125mL flask;</p>
+
 
-
     <p> Melt agarose in microwave until solution is almost boiling, stirring every 15-20 seconds (should be around 2 minutes);</p>
+
<h2>Protocol</h2>
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     <p> Allow agarose to cool (do not let it cool to the point where it is hard);</p>
+
<ol>
-
     <p> Add 5 uL of Red Safe to the cooling agarose;</p>
+
     <li>Measure out 100mL of 1x TAE buffer;</li>
-
     <p> Assemble the gel pouring apparatus by inserting gate into slots;</p>
+
     <li>Transfer buffer to 125 mL flask;</li>
-
     <p> Allow gel to cool until flask can be handled comfortably;</p>
+
     <li>Weigh out enough agarose to make a 1% gel (in our case 1.0 g of agarose was the right amount);</li>
-
     <p> Place comb in the gel rig;</p>
+
     <li> Transfer agarose to 125mL flask;</li>
-
     <p> Pour agarose into gel tray;</p>
+
     <li> Melt agarose in microwave until solution is almost boiling, stirring every 15-20 seconds (should be around 2 minutes);</li>
-
     <p> Allow to solidify. While the gel is solidifying prepare the samples. Add your sample and 1 uL 10x Loading Dye, 4 uL of DNA and 5 uL of water;</p>
+
     <li> Allow agarose to cool (do not let it cool to the point where it is hard);</li>
-
     <p> Pour 1X TAE over gel so that gel is covered by a 3-5mm buffer;</p>
+
     <li> Add 5 uL of Red Safe to the cooling agarose;</li>
-
     <p> Load samples into lane (Don't forget to load a 1kb+ ladder into one of the lanes);</p>
+
     <li> Assemble the gel pouring apparatus by inserting gate into slots;</li>
-
     <p> Hook electrodes to gel apparatus;</p>
+
     <li> Allow gel to cool until flask can be handled comfortably;</li>
-
     <p> Run the apparatus at 100V for 30 - 45 minutes (make sure to watch that the dye does not run off the gel);</p>
+
     <li> Place comb in the gel rig;</li>
-
     <p> Visualize the gel and record the results.</p>
+
     <li> Pour agarose into gel tray;</li>
 +
     <li> Allow to solidify. While the gel is solidifying prepare the samples. Add your sample and 1 uL 10x Loading Dye, 4 uL of DNA and 5 uL of water;</li>
 +
     <li> Pour 1X TAE over gel so that gel is covered by a 3-5mm buffer;</li>
 +
     <li> Load samples into lane (Don't forget to load a 1kb+ ladder into one of the lanes);</li>
 +
     <li> Hook electrodes to gel apparatus;</li>
 +
     <li> Run the apparatus at 100V for 30 - 45 minutes (make sure to watch that the dye does not run off the gel);</li>
 +
     <li> Visualize the gel and record the results.</li>
 +
  </ol>
</section>
</section>
</html>
</html>

Revision as of 04:37, 16 September 2013

Agarose Gel Electrophoresis

TITLE: Agarose Gel Electrophoresis

Reagents and Materials

  • 1X TA
  • Graduated Cylinder
  • 125 mL flask
  • Agarose
  • Gel Pouring Tray
  • Tape
  • Gel rig
  • Red Safe

Protocol

  1. Measure out 100mL of 1x TAE buffer;
  2. Transfer buffer to 125 mL flask;
  3. Weigh out enough agarose to make a 1% gel (in our case 1.0 g of agarose was the right amount);
  4. Transfer agarose to 125mL flask;
  5. Melt agarose in microwave until solution is almost boiling, stirring every 15-20 seconds (should be around 2 minutes);
  6. Allow agarose to cool (do not let it cool to the point where it is hard);
  7. Add 5 uL of Red Safe to the cooling agarose;
  8. Assemble the gel pouring apparatus by inserting gate into slots;
  9. Allow gel to cool until flask can be handled comfortably;
  10. Place comb in the gel rig;
  11. Pour agarose into gel tray;
  12. Allow to solidify. While the gel is solidifying prepare the samples. Add your sample and 1 uL 10x Loading Dye, 4 uL of DNA and 5 uL of water;
  13. Pour 1X TAE over gel so that gel is covered by a 3-5mm buffer;
  14. Load samples into lane (Don't forget to load a 1kb+ ladder into one of the lanes);
  15. Hook electrodes to gel apparatus;
  16. Run the apparatus at 100V for 30 - 45 minutes (make sure to watch that the dye does not run off the gel);
  17. Visualize the gel and record the results.

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