C.B.S. Scientific DGGEK-4801 - Manual (Page 14)

26 27 www.cbsscientific.com Cipher DGGE Instructions 2/28/14
4 2 Buffer Cycling Connections-continued
Buffer Cycling Connections – Dual Cassettes
SECTION 5
Running Conditions
5 1 Running the Gels
1. Load samples at 1:1 with neutral dye. Load 5-10 ug Genomic DNA/well or 1-2 ug
cloned (B-globin)/well. Determine concentration by O.D. 260.
2. Attach black power leads to cassettes as shown in Fig. 5-1.
3 Close lid to engage safety interlock Turn on power supply to 150V (40mA)
constant Volts for 5-7 hours.
5 2 Removing the Gels
3. After the run is completed, turn off power supply, disconnect recirculating tubes
and power leads, remove cassettes, remove glass sandwiches from cassettes.
4. Using a wedge plate separator,cat. # WPS-100, pry plates apart and immerse gel/
plate in buffer tray. Stain with Safe Stain or EtBr (.5ug/ml) for 5-10 minutes. Lift float-
ing gel on plate out of tray and rinse with ddH
2
0. Flip over onto saran wrap and peal
off glass plate. View bands on transilluminator. Electro-Blot or photograph.
5 3 Recommended Power
1. 150V (40mA) CV for 5-7 hours
5 4 Disposal of Buffer Using Buffer Siphon Pump
NOTE: Buffer can be used several times before discarding. Determine empirically.
Dispose of solutions in accordance with the safety regulations of your institution.
1. Attach pump assembly to corner of
tank as shown in figure 5.4a.
2. Place the discharge tubing into a
waste carboy located LOWER than
the pump inlet.
3. Begin siphon with half strokes until
liquid starts to flow. Leave shaft fully
extended as shown in figure 5.4b to
allow siphon to operate.
4. To stop siphon, depress handle and
remove pump from tank.
Fig 5-4a
Fig 5-4b
Fig 5-1
Fig 4-2e