C.B.S. Scientific TTGEK-2401-220 - Manual (Page 47)

TTGE Instruction Manual, version 8/30/2011
47
www.cbsscientific.com
3.12 Perpendicular DGGE Gel Casting
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 Fig 3-11
(top) for casting gels 1.0mm and thicker, see Fig. 3-11 (bottom) for casting gels .75mm or thinner).
2. Follow the Gel Wrap instructions 1-5 (pages 36-37), 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.
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
4. Once the agarose has set, insert the gradient maker outflow tube tipped with a 20ga. through
the Gel Wrap and into the “channel” (top), or space created between the horizontal spacer
and the comb (bottom). 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.
Fig. 3-11 Top
Fig. 3-11 Bottom