Agarose Gel Electrophoresis

Reagents and Materials

• 1X TAE buffer
• Graduated cylinder
• 125 mL flask
• Agarose
• Gel pouring tray
• Tape
• Gel rig
• Red Safe

Protocol

1. Measure 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 μL 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 μL 10x Loading Dye, 4 μL of DNA and 5 μL 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