C.B.S. Scientific TTGEK-2401-220 - Manual (Page 49)

TTGE Instruction Manual, version 8/30/2011
49
www.cbsscientific.com
3.13 Perpendicular Gradient Formation-continued
10. Remove comb and quickly transfer gel sandwich to cassette by removing all #2 clamps. LEAVE GEL WRAP IN
PLACE. The Gel Wrap acts as a barrier and prevents perpendicular electrical fields from interfering with outside
lanes. Re-clamp to cassette using #1 clamps/4 per side. Dislodge the bottom section of the Gel Wrap from the
plates to allow buffer contact with the gel. (Fig 3-7, page 43)
11. Immerse cassette into pre-heated buffer tank, and attach recirculating tubes to each upper reservoir (Fig. 3-8, 3-9).
Rinse loading interface of gel with tank buffer to remove non-polymerized acrylamide or excess urea. Rinse each
well with buffer and fill upper reservoir
12. Load samples.
13. Turn on power supply to 150V (40mA/one dual cassette) constant Volts for 5-7 hours.
14. When run is completed, turn off power supply, disconnect recirculating tubes and power leads, remove cassettes,
remove glass sandwiches from cassettes.
15. Using a wedge plate separator,cat. # WPS-100, pry plates apart and immerse gel/plate in buffer tray. Stain with
EtBr (0.5g/ml) for 5-10 minutes. Lift floating gel on plate out of tray and rinse with DD H
2
0. Flip over onto saran
wrap and peal off glass plate. View bands on transilluminator. Electro-Blot or photograph.