C.B.S. Scientific SG Series - Manual (Page 13)

C.B.S. œ Scientific
13
Adjustable Height Sequencer
3.5 Pouring the Gel
1. Always wear gloves, eye protection and protective clothing .
2. Prepare acrylamide gel solution using freshly made stock solutions or commercially
available pre-mix solutions. See section 4.5 Buffers. Most gel formulations allow
approximately 10 minutes before polymerization, so proceed quickly.
Spacer thickness (mm) Minimum volume required (ml)
0.2 50
0.4 65
0.5 90
0.75-1.0 120-180
3. Mix by gently swirling, using a magnetic spin bar on a stirring plate. Do not mix vigorously.
NOTE: There is no reason to degas acrylamide solutions. This will significantly increase the
speed at which the acrylamide polymerizes. Draw up the solution into a 60cc syringe, 25ml
pipette or an Erlenmeyer flask.
4. If using the Capillary Gel Casting method, pour the gel with the gel plate sandwich in a
horizontal position, while tapping the top plate vigorously to disperse the gel solution.
Capillary action will force the gel towards the bottom and sides. Any small bubbles formed
can be removed easily by using the appropriate thickness bubble grabber.
5. If using the Gel Sealing Tape method, maintain the assembled plate sandwich at a 45 angle.
As the gel sandwich fills, gradually lower the top end of the glass plates until they rest almost
horizontally on the bench. The gel mold should be slightly overfilled to ensure complete
polymerization at the top. If a bubble forms while the gel is being poured, raise the glass
plates into a vertical position, and either tip the gel solution away from the bubble or carefully
tap the plate sandwich. Alternatively, bubbles can be removed by insertion of a bubble
grabber of the same thickness as the spacers being used and gently pulling the bubble or
debris out. Once all bubbles have been removed, return the glass plates to their previous
horizontal position.