Hoefer SE615 User Manual
user manual
Hoefer SE615
and SE675
Multiple gel casters
um SE615-IM/Rev.C0/08-12
Page finder
Safety warnings and precautions ...........................viii
Introduction .......................................................... 1
Setting up the caster ............................................. 5
Casting with divider plates .....................................7
Pouring standard homogeneous gels ........................9
Pouring standard gradient gels ..............................11
Pouring stacking gels ...........................................15
Removing polymerized gels ...................................17
Troubleshooting ...................................................18
Care and maintenance .........................................20
Appendix A: Gel identification numbers ................. 21
Appendix B: References .......................................21
Ordering information ............................................ 22
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Important! Most organic solvents
including methanol, isobutanol,
isopropanol, and even n-butanol
will craze or cloud the acrylic
plastic with prolonged exposure.
Only use small amounts to
overlay the gels.
Safety warnings and precautions
• Acrylamide is a neurotoxin.
• Wear gloves when handling acrylamide or
polyacrylamide.
• Wear a dust mask when weighing acrylamide
or preparing acrylamide solutions.
Protecting your equipment
To keep your instrument in excellent condition,
please take the following important steps:
• Use water-saturated n-butanol for overlayering
resolution gels during polymerization.
• Avoid using small metal spatulas to separate
the plates. Spatulas may chip the edges of glass
plates and prevent them from sealing.
• After casting, you may need to use a wedge
to separate one glass-and-gel sandwich from
another. Slip a Wonder Wedge in between
adjacent sandwiches to separate them.
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Introduction
Gel sizes
The Hoefer® SE615 and SE675 Multiple Gel
Casters are designed for casting vertical slab
acrylamide gels of homogenous or gradient
concentration.
Use the casters with 16 × 18 cm glass plates and
2 cm-wide spacers to cast 16 × 14 cm gels. Use
1-cm wide spacers to cast 16 × 16 cm gels for
two-dimensional electrophoresis. The resulting
gels are compatible with Hoefer SE400, SE600
and SE600 Chroma Vertical Slab Gel Electrophoresis Units.
Homogeneous and gradient gels
The SE615 and SE675 can be used for casting
both homogeneous and gradient gels. Gradient
gels are cast through the port at the bottom of
the caster. When casting homogeneous gels, the
results are most consistent when the gels are cast
from the bottom. The V-shaped region at the
bottom of the casting unit allows the gel solution to rise evenly, producing identical gels.
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Standard casting
In the standard casting procedure, a gel is cast
between two glass plates held apart by a pair of
spacers. (See Fig 2 on page 5.) The result can be
pictured as a “sandwich,” with the gel between
two glass plates. When you set up a vertical slab
gel electrophoresis unit, you place the glass and
gel sandwich in the electrophoresis unit.
The maximum number of gels you can cast
simultaneously using the standard casting procedure is shown in Table 1.
Table 1. Maximum number of
standard gels cast
0.75-mm 1.0-mm 1.5-mm
spacers spacers spacers
SE615 11 10 10
SE675 4 4 4
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Casting with divider plates
You may find it useful to double the number of
gels run simultaneously in one electrophoresis
unit. This can be accomplished by casting a
“club sandwich.” A club sandwich contains a
notched glass divider plate and a second pair
of spacers between the standard plates, and
thus forms two layers of gel between three glass
plates (see Fig 3 on page 8.) Divider plates are
available for the SE400 and SE600.
The maximum number of gels you can cast
simultaneously, using divider plates (two gels
per sandwich), is shown in Table 2.
Table 2. Maximum number of gels
cast with divider plates
0.75-mm 1.0-mm 1.5-mm
spacers spacers spacers
SE615 14 14 12
SE675 6 6 4
See Fig 1 on page 4 to identify the components
of the multiple gel casters. For information on
parts, accessories and related equipment, see
page 22.
Pouring second-dimension slab gels
When preparing second-dimension slab gels in
the multiple casters, it is important to pour the
slabs to the correct height.
If you are using IPG strips, you do not need a
stacking gel. Pour the second-dimension gel to
0.5–1.0 cm below the top of the sandwich for
standard casting, or 1 cm below the notch if
you are casting with divider plates. This allows
ample room to hold the IPG strips between the
two glass plates. Use 1.0- or 1.5-mm spacers
with IPG strips.
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1
5 6
2
3 4
Fig 1. Components of the SE615
and SE675 Multiple Gel Casters.
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8
9
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8
1
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SE615 and SE675 components
no. description
1 Caster body, includes faceplate
2 Spring clamp (4)
3 Foam cord gasket
4 Silicone displacement block*
5 Acrylic spacer block
6 Comb
7 Polycarbonate filler sheet
8 Glass plate
9 Spacer
10 Inlet port
11 Nylon thumb screw
* A small silicone displacement block is also available for the SE675.
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Setting up the caster
Fig 2. Building a
glass sandwich for
standard casting.
glass plate
glass plate
wax paper or
polycarbonate sheet
Note: Do not block the red
inlet port at the bottom of
the faceplate.
caster
spacer
one sandwich
Preparing the caster
1
If the faceplate is attached to the caster, remove it.
Remove the four side clamps, loosen the two thumb
screws, and slide off the faceplate.
2
Remove the foam cord gasket. Apply a light coat of
Gel Seal lubricant and replace the gasket.
3
When casting gradient gels, attach the tubing from the
gradient maker to the inlet port at the bottom of the
faceplate. Homogeneous gels may also be cast through
this port. If casting homogeneous gels from the top,
insert the appropriate displacement block in the open,
V-shaped region at the bottom of the chamber.
4
Place the open caster in a horizontal position with its
back on the bench top.
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Building the stack
Note: To make separation of
polymerized gel sandwiches
easier, place wax paper sheets
in the caster between each set
of glass sandwiches.
Note: When analyzing a large
number of gels, you may
find it useful to include gel
identification numbers into the
polymerized gel.
See the appendix on page 21
for directions on making gel
identification tags. Lightly wet
and place the tags in the lower
corner of the gel sandwich
during stack assembly.
Note: If you use polycarbonate
sheets instead of wax paper
between each sandwich, the
caster holds fewer sandwiches.
1
Place a polycarbonate sheet in the caster so that 1/3
of the sheet extends out of the top.
You can use this sheet as a lever when inserting filler
sheets after all sandwiches are in place. All remaining
components fit flush against the bottom and sides.
2
Place a sheet of wax paper on the polycarbonate
sheet. Build the first sandwich into the caster.
3
Place a sheet of wax paper on top of the sandwich
and build the next sandwich.
4
Repeat step 3, alternating wax paper and gel sandwiches. Over the final sandwich, lay a polycarbonate
sheet instead of wax paper so that the gel solution
level is visible through the faceplate.
5
Place the faceplate on the caster and make sure the
stack fits snugly, about 0.5 mm above the edge of the
caster. If necessary, adjust the number of extra polycarbonate sheets, acrylic spacer blocks, glass plates
and sandwiches to obtain a snug fit.
6
Make sure the spacers are straight and along the
edges of the glass plates. Use the Spacer Mate to
correct the alignment if necessary.
If necessary, take the faceplate off and make sure all
edges of the stack components are flush. Press on
those that are sticking out until all edges are flush
and replace the faceplate on the caster.
7
Secure the faceplate with the four spring clamps and
tighten the bottom thumb screws.
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