"
Team:Calgary/Sandbox/Notebook/Protocols/AgaroseGelElectrophoresis
From 2013.igem.org
(Difference between revisions)
Azzucoloto (Talk | contribs) |
|||
| (One intermediate revision not shown) | |||
| Line 3: | Line 3: | ||
<section id="Content"> | <section id="Content"> | ||
<h1>Agarose Gel Electrophoresis</h1> | <h1>Agarose Gel Electrophoresis</h1> | ||
| - | |||
| - | |||
<h2>Reagents and Materials<h2> | <h2>Reagents and Materials<h2> | ||
<ul> | <ul> | ||
| - | <li>1X | + | <li>1X TAE Buffer</li> |
<li>Graduated Cylinder</li> | <li>Graduated Cylinder</li> | ||
<li>125 mL flask</li> | <li>125 mL flask</li> | ||
| Line 19: | Line 17: | ||
<h2>Protocol</h2> | <h2>Protocol</h2> | ||
<ol> | <ol> | ||
| - | <li>Measure out 100mL of 1x TAE buffer | + | <li>Measure out 100mL of 1x TAE buffer</li> |
| - | <li>Transfer buffer to 125 mL flask | + | <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>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>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>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>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>Add 5 uL of Red Safe to the cooling agarose</li> |
| - | <li>Assemble the gel pouring apparatus by inserting gate into slots | + | <li>Assemble the gel pouring apparatus by inserting gate into slots</li> |
| - | <li>Allow gel to cool until flask can be handled comfortably | + | <li>Allow gel to cool until flask can be handled comfortably</li> |
| - | <li>Place comb in the gel rig | + | <li>Place comb in the gel rig</li> |
| - | <li>Pour agarose into gel tray | + | <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>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>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>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>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>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>Visualize the gel and record the results</li> |
</ol> | </ol> | ||
</section> | </section> | ||
</html> | </html> | ||
Latest revision as of 23:27, 17 September 2013
Agarose Gel Electrophoresis
Reagents and Materials
- 1X TAE Buffer
- 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 TAE Buffer
- 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
