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Team:Calgary/Sandbox/Notebook/Protocols/AgaroseGelElectrophoresis
From 2013.igem.org
(Difference between revisions)
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<h2>Reagents and Materials<h2> | <h2>Reagents and Materials<h2> | ||
<ul> | <ul> | ||
| - | + | <li>1X TA</li> | |
| - | + | <li>Graduated Cylinder</li> | |
| - | + | <li>125 mL flask</li> | |
| - | + | <li>Agarose</li> | |
| - | + | <li>Gel Pouring Tray</li> | |
| - | + | <li>Tape</li> | |
| - | + | <li>Gel rig</li> | |
| - | + | <li>Red Safe</li> | |
</ul> | </ul> | ||
| - | |||
<h2>Protocol</h2> | <h2>Protocol</h2> | ||
<ol> | <ol> | ||
| - | + | <li>Measure out 100mL of 1x TAE buffer;</li> | |
| - | + | <li>Transfer buffer to 125 mL flask;</li> | |
| - | + | <li>Weigh out enough agarose to make a 1% gel (in our case 1.0 g of agarose was the right amount);</li> | |
| - | + | <li>Transfer agarose to 125mL flask;</li> | |
| - | + | <li>Melt agarose in microwave until solution is almost boiling, stirring every 15-20 seconds (should be around 2 minutes);</li> | |
| - | + | <li>Allow agarose to cool (do not let it cool to the point where it is hard);</li> | |
| - | + | <li>Add 5 uL of Red Safe to the cooling agarose;</li> | |
| - | + | <li>Assemble the gel pouring apparatus by inserting gate into slots;</li> | |
| - | + | <li>Allow gel to cool until flask can be handled comfortably;</li> | |
| - | + | <li>Place comb in the gel rig;</li> | |
| - | + | <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 04:48, 16 September 2013
Agarose Gel Electrophoresis
TITLE: Agarose Gel ElectrophoresisReagents and Materials
- 1X TA
- Graduated Cylinder
- 125 mL flask
- Agarose
- Gel Pouring Tray
- Tape
- Gel rig
- Red Safe
Protocol
- Measure out 100mL of 1x TAE buffer;
- Transfer buffer to 125 mL flask;
- Weigh out enough agarose to make a 1% gel (in our case 1.0 g of agarose was the right amount);
- Transfer agarose to 125mL flask;
- Melt agarose in microwave until solution is almost boiling, stirring every 15-20 seconds (should be around 2 minutes);
- Allow agarose to cool (do not let it cool to the point where it is hard);
- Add 5 uL of Red Safe to the cooling agarose;
- Assemble the gel pouring apparatus by inserting gate into slots;
- Allow gel to cool until flask can be handled comfortably;
- Place comb in the gel rig;
- Pour agarose into gel tray;
- 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;
- Pour 1X TAE over gel so that gel is covered by a 3-5mm buffer;
- Load samples into lane (Don't forget to load a 1kb+ ladder into one of the lanes);
- Hook electrodes to gel apparatus;
- Run the apparatus at 100V for 30 - 45 minutes (make sure to watch that the dye does not run off the gel);
- Visualize the gel and record the results.
- 1X TA
- Graduated Cylinder
- 125 mL flask
- Agarose
- Gel Pouring Tray
- Tape
- Gel rig
- Red Safe
Protocol
- Measure out 100mL of 1x TAE buffer;
- Transfer buffer to 125 mL flask;
- Weigh out enough agarose to make a 1% gel (in our case 1.0 g of agarose was the right amount);
- Transfer agarose to 125mL flask;
- Melt agarose in microwave until solution is almost boiling, stirring every 15-20 seconds (should be around 2 minutes);
- Allow agarose to cool (do not let it cool to the point where it is hard);
- Add 5 uL of Red Safe to the cooling agarose;
- Assemble the gel pouring apparatus by inserting gate into slots;
- Allow gel to cool until flask can be handled comfortably;
- Place comb in the gel rig;
- Pour agarose into gel tray;
- 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;
- Pour 1X TAE over gel so that gel is covered by a 3-5mm buffer;
- Load samples into lane (Don't forget to load a 1kb+ ladder into one of the lanes);
- Hook electrodes to gel apparatus;
- Run the apparatus at 100V for 30 - 45 minutes (make sure to watch that the dye does not run off the gel);
- Visualize the gel and record the results.
