Running Electrophoresis Media - GE PhastSystem User Manual

5. Operation
5.4 Running
electrophoresis
media
46
The procedure for running PhastGel electrophoresis media is essentially
the same for both native and SDS techniques. The only operational
difference is the use of PhastGel native or SDS buffer strips and sample
preparation. The general procedure for running PhastGel electrophore-
sis media is described below. (See Separation Technique Files No.110,
111, 112, 120, 121 and 130 for running conditions and more specific
information for each technique.)
Preparing the gel compartment
1. Switch the system on and set the standby temperature to the
temperature of the first step in the method you plan to run first.
(For details see Using the Keyboard, section 5.1.)
2. Adjust the electrode assembly to the high position by pressing up
on both red eccentric levers until they click into place.
3. Raise the electrode assembly to the vertical position. Remove the
IEF gel cover.
4. Wipe off the separation bed with a moist, lint-free cloth to
remove dust or particles. It is also advisable to wipe off the
electrodes, but this must be done gently and the cloth must not
leave dust particles. Avoid touching the electrodes with your
fingers; finger proteins may distort the results.
Note: In order to obtain the best results possible we
recommend frequent cleaning of the electrodes. Even
minor amounts of deposited impurities have been shown
to sometimes affect the resolution and band pattern.
Positioning the gels
1. Place a drop of water or insulating fluid (approximately 60-75 µl)
onto the middle of the gel area(s) outlined by the red lines on the
separation bed.
2. Take one or two gels from the refrigerator. Use a pair of scissors
to cut the package along three sides. Remove the gel from its
package with a pair of forceps; use the plastic tab of the gel
backing as a handle.
The thin plastic film on the gel surface protects the gel from
contaminants and from drying, and should be left on for now.
3. Use a waterproof pen to mark the underside of the gel for
identification. You might have to wipe the back of the gel first.
4. Place the gel on a hard surface and bend the plastic tab up using
the forceps. (This makes it easy to position and remove the gel
from the bed.) Lower the gel onto one of the gel areas so that a
film of liquid, free from air bubbles, forms between the gel
support and separation bed. Remove any air bubbles by sliding
the gel around.