Gene Expression and Proteomics

Gel Electrophoresis

Protocol for Gel Electrophoresis

1. In a flask containing stirring bar, add 0.9 g of agarose (Molecular biology grade).
2. Add approximately 150 mL of 1X TAE Buffer.
3. Microwave for 1 minute. Remove and swirl the flask. Microwave again for 1 minute. Check to make sure all particles of agarose have fully dissolved.
4. On a hot plate, cool down the flask. Do not turn the heat on; turn the stirrer on. Set the timer for seven minutes.
5. Add 7 uL of EtBr solution to the flask.
6. Place two combs on casting platform. Pour solution into the platform. Rinse flask with water.
7. Allow the agarose to set for approximately 20 minutes.
8. Place the casting platform into the gel apparatus. Add more 1X TAE buffer to the tank, if needed (Buffer should cover the wells completely).
9. Wash wells with 1ml pipet, to remove the liquid agarose from wells.
10. Add 5x loading dye to the sample (use about 2 uL per 10 uL sample). Mix well and load into wells.
11. Run for approximately 35 minutes at 80 V (let the front dye move to the middle of gel). Check periodically.
12. Remove the gel and observe under UV light box. Take a picture.