Manual

Cipher DGGE Instructions 2/28/14

3.2 Gel Casting Techniques- continued

B. Vertical gradient gel casting using GM-40 gradient maker and gravity ow.

1. Place the GM-40, gradient maker, on an elevated magnetic stirrer with a small

“ea” stir bar in the cylinder (C-2) closest to the outlet. The gradient maker

should be tted with a leur valve, (V-2) and a 20ga needle with attached tubing

to deliver acrylamide to the gel plate sandwich as shown in gure (3.5). With

the height differential between the leur 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 ow rate. Flow can

be regulated by turning leur valve (V-2). Flow rate appropriate for each gradient

should be determined empirically.

Fig. 3-5

C-1

C-2

V-1

V-2

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