Hoefer SUB Series - Manual (Page 28)

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p20
Gel Pouring
SUB6, SUB10 and SUB13
Units with removable gel casting trays and casting
within the tank.
1
Make sure that the silicone rubber gasket is in
position, i.e. pushed evenly into the groove at the
ends of the gel casting tray.
2 Carefully place the casting tray sideways into the
gel unit so that the gaskets form a seal with the
sides of the running unit.
3 Push the casting tray down onto the gel platform.
4 If the casting tray is too tight, remove the
silicone gaskets and replace it in the end groves
stretching it a little.
5 If the casting tray is too loose, remove the
silicone gaskets and replace them in the end
grooves compressing it a little.
6 Place the gel unit on a level surface.
7 Position the required comb(s) into the slot(s) in
the gel casting tray.
8 Prepare 22.5, 60 or 100 mls of the desired
% agarose for units SUB6, SUB10 or SUB13
respectively. This will give a gel depth of 5 mm.
For a 10 mm deep gel, prepare 45, 125 or
200 mls respectively.
9 Using a 1 ml disposable pipette, seal the silicone
seal with some of the cooled agarose.
10 Pour in the agarose smoothly so as not to
generate bubbles. If bubbles do occur, these can
be smoothed to the side of the gel and dispersed
using a clean gloved hand.
11 Allow the agarose to set, ensuring that the gel
remains undisturbed.
12 Carefully remove the gel casting tray from the
running unit. Either remove the end silicone
rubber gaskets, or leave them in place.
13 Place the gel casting tray in the running position
such that the wells are nearest to the black
electrode (cathode).
14 Fill the tank with running buffer so that the
buffer just submerges the gel.
15 Carefully remove the comb.
IMPORTANT! Ensure that the
agarose has cooled to between
50 and 60 °C to prevent
apparatus distortion.