Manual

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3.6GelCastingTechniques-continued

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

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 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 ow 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.