"
Team:Calgary/Notebook/Protocols/AgaroseGelElectrophoresis
From 2013.igem.org
(Difference between revisions)
(Created page with "<html> <div id="Banner"><h1>Agarose Gel Electrophoresis</h1></div> </html> {{Team:Calgary/ContentPage}} <html> <section id="Content"> <h1>Agarose Gel Electrophoresis</h1> <h2>Re...") |
Sharonfeng1 (Talk | contribs) |
||
| (One intermediate revision not shown) | |||
| Line 9: | Line 9: | ||
<h2>Reagents and Materials<h2> | <h2>Reagents and Materials<h2> | ||
<ul> | <ul> | ||
| - | <li>1X TAE | + | <li>1X TAE buffer</li> |
| - | <li>Graduated | + | <li>Graduated cylinder</li> |
<li>125 mL flask</li> | <li>125 mL flask</li> | ||
<li>Agarose</li> | <li>Agarose</li> | ||
| - | <li>Gel | + | <li>Gel pouring tray</li> |
<li>Tape</li> | <li>Tape</li> | ||
<li>Gel rig</li> | <li>Gel rig</li> | ||
| Line 20: | Line 20: | ||
<h2>Protocol</h2> | <h2>Protocol</h2> | ||
<ol> | <ol> | ||
| - | <li>Measure | + | <li>Measure 100mL of 1x TAE buffer</li> |
<li>Transfer buffer to 125 mL flask</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>Weigh out enough agarose to make a 1% gel (in our case 1.0 g of agarose was the right amount)</li> | ||
| Line 26: | Line 26: | ||
<li>Melt agarose in microwave until solution is almost boiling, stirring every 15-20 seconds (should be around 2 minutes)</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>Allow agarose to cool (do not let it cool to the point where it is hard)</li> | ||
| - | <li>Add 5 | + | <li>Add 5 μL of Red Safe to the cooling agarose</li> |
<li>Assemble the gel pouring apparatus by inserting gate into slots</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>Allow gel to cool until flask can be handled comfortably</li> | ||
<li>Place comb in the gel rig</li> | <li>Place comb in the gel rig</li> | ||
<li>Pour agarose into gel tray</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 | + | <li>Allow to solidify. While the gel is solidifying prepare the samples. Add your sample and 1 μL 10x Loading Dye, 4 μL of DNA and 5 μL of water</li> |
<li>Pour 1X TAE over gel so that gel is covered by a 3-5mm buffer</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>Load samples into lane (Don't forget to load a 1kb+ ladder into one of the lanes)</li> | ||
Latest revision as of 00:15, 28 September 2013
Agarose Gel Electrophoresis
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 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 μL 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 μL 10x Loading Dye, 4 μL of DNA and 5 μL 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 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 μL 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 μL 10x Loading Dye, 4 μL of DNA and 5 μL 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
