Gel Electrophoresis Procedure
Casting a 0.8% Agarose Gel
1.
Use a 250 mL flask to prepare the gel solution. To the flask add 0.24 g of
agarose, 0.6 mL concentrated (50x) buffer, and 29.4 mL distilled water. You will
have a total volume of 30 mL. Swirl the mixture to disperse any clumps of
agarose powder.
2.
Us a marking pen to indicate the level of the solution volume on the outside of the
flask.
3.
Cover the flask with plastic wrap to minimize evaporation. Place the flask in a
microwave oven and heat on high for 1 minute. Swirl the mixture and continue
heating on high in bursts of about 25 seconds until all the agarose has dissolved.
4.
Coll the agarose solution to 55 oC with careful swirling to promote even
dissipation of heat. Make sure that the level of the liquid is the same as before
heating. If not, add distilled water to bring the volume up to the original volume.
5.
Pull up the dams on the ends of a 7 x 7 gel bed. Place the bed on a level surface,
add a six tooth comb and pour the solution into the bed.
6.
Let the gel completely solidify. It will be firm and cool to the touch. This will
take approximately 20 minutes.
Preparing the Gel of Electrophoresis:
1.
Drop the gates and carefully remove the comb by slowly pulling it straight up.
You do not want to tear the sample wells.
2.
Place the gel bed into the electrophoresis chamber