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3.9 Perpendicular Gel Casting
NOTE:
Prior to casting, mark the notched glass plate to designate which side is the ‘inside’ and
‘outside’. The same side of the notched plate should always sit against the reservoir gasket.
1. For perpendicular gel casting, locate the spacer which has a channel machined into it on
one end which will be referred to as the "channel spacer". The gel sandwich will be cast
on its side, with the channel spacer on top and the channel further away from the single
well comb (see figure 3-11 top for casting gels 1.0mm and thicker, see figure 3-11 bottom
for casting gels .75mm or thinner).
2. Follow the Gel Wrap instructions 1-5 (pages 12-13),
making sure to use the channel
spacer
.
3. Insert single well comb into glass notch. A small amount of agarose is needed in two
places once the gel sandwich is placed on its side, both places are located on the lower
corner
where the single well comb interfaces the spacer, and on the other side of the
spacer where it interfaces the gel wrap.
4. Once the agarose has set, insert the gradient maker outflow tube tipped with a 20ga.
through the Gel Wrap and into the "channel" (figure 1), or space created between the
horizontal spacer and the comb (figure 2). You may use the same piece of Gel Wrap
many times (20-30) for the perpendicular gels: do not use a piece of Gel Wrap Gasket for
Vertical casting after you have poked a hole in it for perpendicular casting—it will leak.
FIGURE 1
FIGURE 2
1.0mm AND THICKER
.75mm AND THINNER
PERPENDICULAR GEL CASTING
TO GRADIENT MAKER
20 GAUGE NEEDLE
SEAL WITH AGAROSE
HOLE THROUGH SPACER
GEL PLATE
CLAMPS
TO GRADIENT MAKER
20 GAUGE NEEDLE
SEAL WITH AGAROSE
GEL PLATE
CLAMPS
Fig. 3-11 Top
Fig. 3-11 Bottom
Fig. 3-11
