Team:University College London/Protocols/Electrophoresis

Electrophoresis

Preparing the Gel

1. Within a conical flask, add 3ml 50X TAE, 1.5g Agarose, and 150ml RO water

2. Heat for 1 min in microwave. Swirl. Heat again for 30s. If solution is clear stop. Else repeat.

3.Cool solution under running cold water.

4. Add 20ul ethidium bromide (normal concentration of EB solution is 500ug/ul)

5. Pour into a sealed casting tray in a slow steady stream, ensuring there are no bubbles

Running a gel

1. Add 1 part loading buffer to five parts of loading sample

2. Position the gel in the tank and add TAE buffer, enough to cover the gel by several mm

3. Add 5ul of DNA ladder to lane 1

4. Add samples to the remaining wells

5. Run at 100 volts for 1hour and 15 minutes

Imaging the Gel

1. Place gel in GelDoc 2000 chamber

2. Turn GelDoc 2000 chamber on 

3. From computer: Quantity One > Scanner > Gel_Doc_Xr>Manuqal Acquire

4. Alter the exposure/settings to give a clear image.

TAE - Tris-acetate-EDTA EDTA - ethylenediamine tetraacetic acid