Instruction Manual
Gel Multi Caster
6 x (for 6 gels)
12 x (for 12 gels)
3117.1
3118.1
PACKING LIST
Art. No.
3117.1
3118.1
Bottom Gasket
for Top Filling
1
1
Leveling
Bubble
1
1
12 mm
Saver Plate
1
1
5 mm
Saver Plate
3
5
3 mm
Saver Plate
2
3
Separation
Sheets
12
12
Please verify that you received the unit completely and without any damage. Any faults or losses have
to be reported to ROTH immediately. ROTH can not accept responsibility for goods that were sent
back without informing them.
Please take a look at the packing list and check whether all components and accessories are present.
CARE AND MAINTENANCE
Cleaning Gel Multi Caster Units
Units are best cleaned using warm water and a mild detergent.
Please note: Water at temperatures above 60 °C can cause damage to the unit and components.
The tank should be thoroughly rinsed with warm water or distilled water to prevent build up of salts.
Vigorous cleaning is not necessary or advised. Air drying is preferably before use.
The units should only be cleaned with the following:
Warm water with a mild concentration of soap or other mild detergent. Compatible detergents include
dishwashing liquid, Hexane and Aliphatic hydrocarbons.The units should not be left in detergents for
more than 30 minutes.
Please note: The units should never come into contact with the following cleaning agents; these will
cause irreversible and accumulative damage:
Acetone, Phenol, Chloroform, Carbon tetrachloride, Methanol, Ethanol, Isopropyl alcohol, Isobutanol
or any other alcohol. Alkalis.
RNase Decontamination
Clean the units with a mild detergent as described above.
Wash with 3% hydrogen peroxide (H2O2) for 10 minutes.
Rinsed with 0.1% DEPC- (diethyl pyrocarbonate) treated distilled water,
Caution: DEPC is a suspected carcinogen. Always take the necessary precautions when using.
RNaseZAP™ (Ambion) can also be used. Please consult the instructions for use with acrylic gel
tanks.
1
OPERATION
A. Top Filling – Standard Single Percentage Gels
Note: Use only glass plate sandwiches consisting of one notched glass plate and one plain glass
plate with bonded spacers.
1.
Use the levelling bubble provided to ensure that the surface is level upon which the caster to be
set up.
2.
Loosen the clamp screws, remove the caster door, and place the open casting chamber on the
bench top so that the centre of the casting chamber is facing upwards.
Please note: Make sure that the white silicone gasket, supplied for top filling, is in the bottom of
the casting chamber. Ensure that the flat surface is pointing upwards. The V-shaped ridge on
the underside of the white silicone gasket should also fit snugly in the V-shaped channel within
the casting chamber.
3.
Insert the glass and spacer plates as follows:
Using the Gel Multi Caster for 6 gels (3117.1) as an example, there should be a separation
sheet at the rear of the casting chamber followed by a glass plate sandwich (1 mm spacer). This
combination of separation sheet and glass plate sandwich is repeated a further five times. Using
the Gel Multi Caster for 12 gels (3118.1) corresponding more plates are necessary.
Then a 5 mm spacer plate is inserted followed by a 3 mm spacer plate and a further two
separation sheets. At this point the glass and spacer plate stack should be flush with the edges
of the casting chamber.
4.
Overlay the caster door and secure it to the casting chamber by tightening the screws. The front
gasket should flatten all the way around which indicates a good seal.
5.
Restore the casting chamber to its upright position without dislodging the glass plates. If, by
tightening the screws, the glass plates have been pushed upwards push them downwards to
ensure that they retain contact with the silicone mat. The spare 5mm spacer plate may be used
to push each glass plate sandwich back into position.
6.
Make sure that the clip provided is used to seal the silicone hose at the front.
7.
Pour pre-prepared acrylamide solution (including APS and TEMED initiators) into the top of the
glass plate sandwich closest to the front of the casting chamber (i.e. in the cavity between the
plain glass plate with bonded spacers and the notched glass plate). This should be performed
evenly and smoothly to allow the gel level to equilibrate.
It is much easier if the required volume has been determined previously and prepared
accordingly (e.g. by filling the assembled gel caster - containing glass plates, spacer plates and
separation sheets - with water and noting the volume required).
If this is not the case then fill the middle plates and look for when the end plates reach the
desired level.
Please note: The purpose of the white silicone mat is not to stop leakage as in a standard
casting base, but to provide even gel equilibration and distribution between the individual gel
sandwiches, in addition to minimizing the volume of acrylamide required. The acrylamide will
migrate by capillary action from the front gel sandwich to the other gel sandwiches in the
caster. This is entirely normal, and necessary to fill all of the sandwiches evenly.
Please note: Once the acrylamide has set the separation sheets may be used to pry apart the
individual gel sandwiches as described at the end of this protocol.
8.
The gel will have to be allowed to set and a suitable overlay (e.g. distilled water) applied if a
stacking layer is to be poured above the resolving layer.
9.
Insert the combs and allow the gels to set.
2
OPERATION
B. Bottom Filling – Gradient Gels
Note: Use only glass plate sandwiches consisting of one notched glass plate and one plain glass
plate with bonded spacers.
1.
Use the levelling bubble provided to ensure that the surface is level upon which the caster to be
set up.
2.
Loosen the clamp screws, remove the caster door, and place the open casting chamber on the
bench top so that the centre of the casting chamber is facing upwards.
Please note: Remove the large bottom white gasket for top-filling, to reveal a V-shaped
channel. This channel is necessary for acrylamide gradient equilibration when the gel is poured.
3.
Insert the glass and spacer plates as follows:
Using the Gel Multi Caster for 6 gels (3117.1) as an example, there should be a separation
sheet at the rear of the casting chamber followed by a glass plate sandwich (1 mm spacer).
This combination of separation sheet and glass plate sandwich is repeated a further five times.
Using the Gel Multi Caster for 12 gels (3118.1) corresponding more plates are necessary. Then
a 5 mm spacer plate is inserted followed by a 3 mm spacer plate and a further two separation
sheets. At this point the glass and spacer plate stack should be flush with the edges of the
casting chamber.
4.
Overlay the caster door and secure it to the casting chamber by tightening the screws. The
Front gasket should flatten all the way around which indicates a good seal.
5.
Restore the casting chamber to its upright position without dislodging the glass plates. If, by
tightening the screws, the glass plates have been pushed upwards push them downwards to
ensure that they retain contact with the silicone mat. The spare 5mm spacer plate may be used
to push each glass plate sandwich back into position.
6.
The first time a certain number of gels of a certain thickness are cast, it is necessary to
determine the required volume of acrylamide so as to ensure a complete gradient. This can be
done by calculating the volume of gel in each plate and multiplying by the number used. There
is also 8ml of gel which will not fill the glass plates. This will fill the bottom gap and is necessary
for even filling of the plates. This should be taken into consideration and added to the lowest
gradient.
7.
Place the Gradient Mixer (N857.1 / N858.1) on a magnetic stirring plate and attach the outlet to
the port on the Gel Multi Caster. Ensure the outlet valve on the Gradient Mixer is closed.
The following steps need to be performed, smoothly sequentially and rapidly to ensure correct and
complete pouring. It is usually necessary to practice these steps beforehand to ensure that the
polymerization time of the gel is not exceeded by the pouring time. If this is the case the initiators can
be adjusted to allow polymerization over a longer time period.
8.
Determine the volume of both high and low concentrations of gel necessary and immediately
prior to inserting, add APS and TEMED. Then quickly pour the solutions into the correct
chambers of the Gradient Mixer as per the manufacturer’s instructions.
9.
Turn on the stirring bar in the mixing chamber of the Gradient Mixer, open the tubing clamp of
the Gradient Mixer and allow the Gel Multi Caster to fill. This may need a peristaltic pump if the
flow is not fast enough.
10.
Insert the combs and allow the gels to set.
3
REMOVAL AND STORAGE OF GELS
Note: Wear rubber gloves while performing the following procedure to prevent accidental exposure to
unpolymerized acrylamide.
1.
Unscrew the clamp screws and remove the front plate.
2.
Remove the gels one at a time front first from the stack. Carefully separate the gel sandwiches
from the separation sheets and any acrylic blocks that were used as space-fillers.
3.
Rinse off the tops of all the gels thoroughly with distilled water.
4.
Store the gels upright in a tightly sealed container. Add a few millilitres of 1X gel buffer (identical
to the buffer in the gel) to the bottom of the container and to the tops of the gels to prevent them
from drying out.
5.
Store tightly sealed at 4 °C.
6.
Clean the entire casting chamber thoroughly with distilled water. Residual acrylamide in the
stopcock valve and inlet port can be removed using a paper clip or a syringe needle.
Carl Roth GmbH + Co. KG
Schoemperlenstraße 3-5
76185 Karlsruhe
Postfach 100121
76231 Karlsruhe
Telefon: +49 (0) 721/ 5606-0
Telefax: +49 (0) 721/ 5606-149
E-mail: [email protected]
Internet: www.carlroth.de
s.t. 03/2017
4
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