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Versión/Version 1, Sept 2017
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Rinse with 0.1% DEPC-(diethyl pyrocarbonate) treated distilled water. Caution!: DEPC
is a suspected carcinogen. Always take the necessary precautions when using.
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RNaseZAP
TM
(Ambion) can also be used. Please consult the instructions for use with
acrylic gel tanks.
4.
Setting up the Horizontal Gel Tanks:-
Instructions for fitting Electrode Cables
1.
Note the position of the lid on the unit. This shows the correct polarity and the correct
orientation of the cables, black is negative and red positive.
2.
Remove the lid from the unit, Note if the lid is not removed, fitting the cables may result
in un-tightening of the gold plug and damage to the electrode.
3.
Screw the cables into the tapped holes as fully as possible to that there is no gap
between the lid and the leading edge of the cable fitting.
4.
Refit the lid.
5.
Gel Preparation
1.
For a standard 0.7% agarose gel, add 0.7 g of agarose to 100 mL of 1x TAE or TBE solution.
The same 1x solution should be used in the tank buffer solution.
2.
Add the agarose powder to a conical flask.
3.
Add the appropriate amount of 1X TAE or TBE solution. To prevent evaporation during the
dissolving steps below, the conical flask should be covered with parafilm.
4.
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 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.
5.
The gel must be cooled to between 50º C and 60ºC before pouring.
6.
Gel Pouring
1.
Put the gel mold onto a level surface and put a fit gel tray into it. In order to prevent gel
leaking, both ends of gel tray must be closely against the gel box.
2.
Place the comb(s) onto the tray.
3.
Pour agarose carefully so as not to generate bubbles.
4.
Leave the gel by itself and wait it to concrete.
