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.