Final Assembly of Apparatus

Final Assembly of Apparatus
1. Lower Top Reservoir around Cooling Unit.
Electrical posts should be on the same side.
2. Disperse bubbles under gels using curved pipet.
3. Slowly add the remaining Upper Tris Buffer to the Top Reservoir.
Minimize turbulence to prevent disrupting samples in wells.
Note level of Tris Running Buffer in Top Reservoir.
4. Place on magnetic stirrer.

Run Gels
1. Attach lid and connect electrical leads.
2. Turn-on power supply with rocker switch on lower left side.
3. Push the Select Mode membrane switch to the toggle to the mA position.
4. Push the Display membrane switch to toggle to Set position.
5. Adjust to 17.5 mA/gel (1.5 mm) using the large knob.
6. Push the the On/Off membrane switch to start run.
7. Record the mA, V and time on the The Gel Run Log.

Post-Run Disassembly
1. Add 200 ml 25% Trichloroacetic Acid (TCA) to a 500 ml graduated cylinder.
2. Add 200 ml RO H2O to the cylinder for 400 ml total.
3. Pour this solution into plastic staining tray and place next to sink.
4. Record final mA, V and time on the The Gel Run Log.
5. Turn off the power supply first with membrane switch then rocker switch.
6. Detach lid and electrical posts.
7. Slide Top Reservoir upward and move to sink.
8. Pour buffer in Top Reservoir into sink.
9. Lay assembly on its side
10. Remove cam levers on top gel set while holding glass plates.
11. Separate glass plates with plastic or wooden spatula.
DO NOT USE METAL.
12. Observe which plate the gel is adhering to and move the glass plate so that the gel is on top.
13. Place the glass plate on a clean level surface
14. Slice off the Upper (stacking) Gel approximately 1-2 mm above the Lower (resolving) gel interface using a single-edged razor blade.
15. Touch a paper towel to the Upper (stacking) Gel in order to remove it.
Discard in HAZARDOUS WASTE container.
16. Rinse lightly with a stream of DI H2O.
Tilt the plate upward so that if the gel were to slide off, it would be blocked by your hand.
17. Invert the glass plate over the staining tray
18. Lower the glass plate into the solution and then lift so that the surface tension will peel the gel from the glass plate.
Use a plastic or wooden spatula to loosen one cornor of the gel if necessary.
19. repeat the same procedure with the other gel(s).