15/11/2018
Page 9
Gel Preparation
The Table below shows the volume of agarose solution required to make the
desired agarose gel for each unit tray size. For a standard 0.7% agarose gel,
add 0.7 grams of agarose to 100 ml of 1x TAE or TBE solution. The same 1X
solution should be used in the tank buffer solution.
Tray
15 x 7 cm
15 x 10 cm
15 x 15 cm
Gel volume for a 5mm thick gel
52.5 mL
75 mL
112.5 mL
1. Add the agarose powder to a conical flask.
2. Add the appropriate amount of 1x TAE or TBE solution from the table
above. To prevent evaporation during the dissolving steps below, the
conical flask should be covered with parafilm.
3. Dissolve the agarose powder by heating the agarose either on a
magnetic hot plate with stirring bar or in a microwave oven. If using the
microwave method, the microwave should be set at around a 400 watt
or medium setting and the flask swirled every minute. The solution
should be heated until all crystals are dissolved. This is best viewed
against a light background. Crystals appear as translucent crystals.
These will interfere with sample migration if not completely dissolved.
The gel must be cooled to between 50°C and 60°C degrees before pouring.
For Real-Time visualisation, mix a compatible DNA stain such as Ethidium
Bromide with your agarose gel in the required proportion. This allows DNA to
be visualised during the run.
Alternatively, runSAFE DNA stain can be used as the sample loading buffer to
enable real-time visualisation.
