Gel Electrophoresis

Gel electrophoresis is a widely used technique for the analysis of nucleic acids and proteins. Most every molecular biology research laboratory routinely uses agarose gel electrophoresis for the preparation and analysis of DNA. We will be using agarose gel electrophoresis to determine the presence and size of PCR products.

Preparing the agarose gel

• Measure 1.25 g Agarose powder and add it to a 500 ml flask • Add 125 ml TAE Buffer to the flask. (the total gel volume well vary depending on the size of the casting tray) • Melt the agarose in a microwave or hot water bath until the solution becomes clear. (if using a microwave, heat the solution for several short intervals - do not let the solution boil for long periods as it may boil out of the flask). • Let the solution cool to about 50-55°C, swirling the flask occasionally to cool evenly. • Seal the ends of the casting tray with two layers of tape. • Place the combs in the gel casting tray. • Pour the melted agarose solution into the casting tray and let cool until it is solid (it should appear milky white). • Carefully pull out the combs and remove the tape. • Place the gel in the electrophoresis chamber. • Add enough TAE Buffer so that there is about 2-3 mm of buffer over the gel.

Preparing the agarose gel 1%:

1. Measure 1 g Agarose powder and add it to a 500 ml flask

2. Add 100 ml TAE Buffer to the flask

3. Melt the agarose in a microwave until the solution become clear (if using a microwave, heat the solution for several short intervals - do not let the solution boil for long periods as it may boil out of the flask).

4. Let the solution cool to about 50-55°C, swirling the flask occasionally to cool evenly

5. Add BET with a final concentration of 200ng/µL

6. Seal the ends of the casting tray with two layers of tape

7. Place the combs in the gel casting tray.

8. Pour the melted agarose solution into the casting tray and let cool until it is solid (it should appear milky white).

9. Carefully pull out the combs and remove the tape

10. Place the gel in the electrophoresis chamber.

11. Add enough TAE Buffer so that there is about 2-3 mm of buffer over the gel.

Put the misted DNA with charged blue diluted 6X