Richard Lab:Agarose Gel Electrophoresis

This protocol is for the typical separation and purification of DNA done in the richard lab. The consensus Electrophoresis protocol should be consulted if deviating from the procedure outlined here.

Procedure

1. Place the gel tray perpendicular in the electrophoresis to create a casting bay.
2. Wipe the tray clean with a Kim-wipe and level the tray using the bubble level.
3. Add the desired amount of agarose to 100ml of 1X TBE in a 300ml flask
4. Microwave the flask on high for 80 seconds. Be sure that it does not boil over.
5. Swirl the microwaved agarose in the flask until the solution becomes clear.
6. Pour the melted solution into the casting bay and insert the comb.
7. Prepare the DNA ladder by combining the following:

• 10ul DNA ladder
• 1ul SYBR green (1:100 Dilution in DMSO)
• 1ul Bromophenol Blue

The amount of agarose to use in your gel depends on the DNA in question. As a rough guide: Agarose (g/100mL) DNA resolution (/kb) 0.5 1-30 0.7 0.8-12 1.0 0.5-10 1.2 0.4-7 1.5 0.2-3