C.B.S. Scientific TTGEK-2401-220 - Manual (Page 38)

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3.6 Gel Casting Techniques- continued
B. Vertical gradient gel casting using GM-40 gradient maker and gravity flow for DGGE.
1. Place the GM-40, gradient maker, on an elevated magnetic stirrer with a small “flea” stir bar in
the cylinder (C-2) closest to the outlet. The gradient maker should be fitted with a luer valve, (V-
2) and a 20ga needle with attached tubing to deliver acrylamide to the gel plate sandwich. (Fig
3-4). With the height differential between the luer valve (V-2) and the top of the gel plate sandwich
between 2-4 inches, it takes approximately 5-9 minutes to cast the gel. Raising the height of the
gradient maker can increase flow rate. Flow can be regulated by turning luer valve (V-2). Flow
rate appropriate for each gradient should be determined empirically.
C-1
C-2
V-1
V-2
Fig. 3-4
2. For TTGE and SSCP single percentage gels, use a pipette or syringe to inject polyacrylamide into
glass plate cassette.
Refer to section 3.7 for Gradient Gel Formula Protocol.
See section 4.3 for running conditions.