VWR Peqlab Cast-It User manual

Instruction Manual – Bedienungsanleitung
Cast-It™ Gel CASTER /
Cast-It™ Gelgießstand

Instruction Manual Cast-It™ Gel CASTER
VWR v0617 E
CONTENT
WARRANTY 1
PACKAGING LIST 1
ASSEMBLING GEL PLATES FOR CASTING 2
BUFFERS AND SOLUTIONS 3
TROUBLESHOOTING 4
TECHNICAL SUPPORT AND ORDERING INFORMATION 5

Instruction Manual Cast-It™ Gel CASTER
VWR v0617 E 1
WARRANTY
VWR guarantees that the system you have received has been thoroughly tested and meets its published
specification.
However, immediately upon arrival, please check carefully that the shipment is complete and has not
been damaged during transit. For missing parts or to report any kind of damage, please contact VWR
(see 'TECHNICAL SUPPORT AND ORDERING INFORMATIONS'). Please retain all packaging materials
until the delivery has been completely checked since this will speed up the return of goods if required
and reduce environmental impact. Any form of returns, replacements or credit notes must be agreed in
advance by VWR.
For the complete range of PerfectBlue
™
electrophoresis and blotting systems, VWR guarantees a
warranty period of 36 months if the products have been used solely according to the instruction
manual, if not agreed differently. No liability is accepted for loss or damage arising from incorrect use.
VWR's liability is limited to the repair or replacement of the unit or refund of the purchase price, at
VWR's discretion. VWR is not liable for any consequential damages. After the warranty period has
expired VWR can offer repairs at low costs.
VWR reserves the right to alter the technical specifications of the PerfectBlue
™
electrophoresis or blotting
systems without prior notice. This will enable us to implement developments as soon as they arise.
PACKAGING LIST
Unless differently agreed or marked on the delivery note the following items are included in shipment
for the Cast-It™ Gel Caster:
Casting base
Center unit with side clamps
2 casting cams
2 gaskets for the center unit (red)
2 gaskets on the casting base (black)
When using the casting base the first time remove the plastic strips from the black gaskets.
Note: Glass plates, spacers and combs are not included in the shipment and should be ordered
separately if needed!

Instruction Manual Cast-It™ Gel CASTER
VWR v0617 E 2
ASSEMBLING GEL PLATES FOR CASTING
Picture 1: Invert casting base.
Picture 2: Place gel assembly.
Picture 3: Turn handles.
See Table 1 for volumes of acrylamide mixture needed. Pour gel solution slowly to avoid any
bubbles. When gel is poured place comb at top to create wells.
Remove gels from the Cast-It™ center unit and place them into the electrophoresis chamber.
Table 1:
Unit
Twin S
Twin M
Twin ExW S
Twin L
0.8 mm spacer
8 ml
18 ml
16 ml
32 ml
1.5 mm spacer
15 ml
34 ml
30 ml
60 ml

The center unit of the Cast
-
It™ is attached to the casting
base with safety cams and slot arrangement.
For casting gels remove the center unit from the base.
Invert casting base (picture 1).
Place the center of the Cast-It™ on the inverted casting
base. The notches of the center unit need to lock into
the four acrylic squares on the underside of the casting
base. Arrange a gel assembly including a notched
glass plate, side spacers and a blank glass plate. Place
the assembly onto one side of the center unit. Slide the
side clamps over the assembly and tighten the screws
(picture 2). DO NOT OVERTIGHTEN. Place a second
gel assembly on the other side. If you only want to cast
one gel, place a blank glass plate or a blocking plate,
on the other side. The bottom of the glass plates needs
to sit flat on the acrylic squares of the inverted casting
base to obtain a good seal. Remove the center unit and
turn around the casting base.
Wet silicone mats with e.g. water or buffer to prolong
their life span.
Place the center unit onto the casting base. Insert the
casting cams (with handles pointing up) into the holes
in the center unit. Simultaneously, turn cams one half
turn to tighten the assembly onto the gasket of the
casting base.
Once the glass plates are secured into the casting base
the gel may be poured. (An initial leak test using a
small amount of water is recommended - add 2 - 3 ml
of water and let stand for 2 minutes. If no leakage is
visible, remove water and proceed with gel casting. If
leakage occurs, take assembly apart and try again.)

Instruction Manual Cast-It™ Gel CASTER
VWR v0617 E 3
A few tips about caring for your system:
Clean all VWR acrylic systems with warm water and a mild detergent. Do not autoclave, bake or
microwave your unit. Temperatures over 50 °C can cause damage to the acrylic.
Always wet silicone mats with e.g. water or buffer before placing the center unit.
WARNING! DO NOT USE ETHANOL OR OTHER ORGANIC SOLVENTS TO CLEAN VWR PER-
FECTBLUE™ PRODUCTS. Organic solvents cause acrylic to 'craze' or crack.
BUFFERS AND SOLUTIONS
1 x Tris-Glycine electrophoresis buffer
25 mM Tris
250 mM glycine (electrophoresis grade) pH 8.3
0.1 % SDS
Prepare a 5 x stock solution of electrophoresis buffer by dissolving 15.1 g Tris base and 94 g glycine in
900 ml of deionized H
2
O. Add 50 ml of a 10 % stock solution of SDS (electrophoresis grade) and
adjust the volume to 1000 ml with H
2
O.
Resolving Gels for Tris-Glycine SDS-PAGE
6 % 8 % 10 % 12 % 15 %
H
2
O 5.3 4.6 4.0 3.3 2.3
30 % acrylamide mix 2.0 2.7 3.3 4.0 5.0
1.5 mM Tris (pH 8.8) 2.5 2.5 2.5 2.5 2.5
10 % SDS 0.1 0.1 0.1 0.1 0.1
10 % ammonium persulfate 0.1 0.1 0.1 0.1 0.1
TEMED 0.008 0.006 0.004 0.004 0.004
All amounts in ml per 10 ml gel solution
5 % Stacking Gels for Tris-Glycine SDS-PAGE
1 ml 2 ml 3 ml 5 ml 10 ml
H
2
O 0.68 1.4 2.1 3.4 6.8
30 % acrylamide mix 0.17 0.33 0.5 0.83 1.7
1.0 mM Tris (pH 6.8) 0.13 0.25 0.38 0.63 1.25
10 % SDS 0.01 0.02 0.03 0.05 0.1
10 % ammonium persulfate 0.01 0.02 0.03 0.05 0.1
TEMED 0.001 0.002 0.003 0.005 0.01
All amounts in ml

Instruction Manual Cast-It™ Gel CASTER
VWR v0617 E 4
TROUBLESHOOTING
Below are some possible solutions to potential problems. If these suggestions are unsuccessful, do not
hesitate to contact us (see 'TECHNICAL SUPPORT AND ORDERING INFORMATIONS').
Problem Cause Solution
Leakage of gel
solution
Glass plates and spacers have
not been aligned correctly
For correct alignment of glass plate sandwich it is necessary
to place the upper buffer chamber on top of the inverted
casting base. The bottom of the glass plates and the spacers
need to sit flat on the acrylic squares.
Glass plates or spacers are
dirty
Clean thoroughly.
Uneven edges of glass plates Only use glass plates with polished edges.
Silicone gaskets of casting
base are dirty
Clean thoroughly with a mild detergent and rinse with
distilled water, dry without leaving smearing, i.e. air dry with
a non-smearing paper or washleather.
Silicone gaskets of casting
base are brittle
Replace silicone gaskets which can be easily removed. New
gaskets will stick due to their own adherence. Also, wet
silicone mats with e.g. water or buffer before placing the
center unit onto the casting base to prolong their life span.
Skewed bands Gel has not polymerized
properly at wells.
De-gas gel solution before casting and increase APS and
TEMED concentrations. The comb can be wiped with TEMED
just prior to casting to improve polymerization.
Resolving gel is uneven at top.
Overlay gel carefully using water saturated n-butanol and
make sure casting stand is level.
The upper buffer chamber is
leaking either through the gel
or along the side spacers.
Check gel to make sure that it is a solid slab inside the glass
and check the setup of the apparatus to ensure a seal with
the gasket. Always use combs and spacers of the same
thickness.
'Frowning' of
outside lanes
Leakage of buffer along sides
or along
spacers inside the gel
assembly.
Do not move spacer after polymerisation of gel. Make sure
that the glass plates are pressed firmly against the gasket of
the UBC.
Silicone mats wear
out quickly
Sticky surface of the mats
adheres too strongly to the
glass plates. Upon removal of
the center unit silicone mats
are torn apart.
Wet the silicone mats with e.g. water or buffer before placing
UBC onto the casting base to prolong their life span.

Instruction Manual Cast-It™ Gel CASTER
VWR v0617 E 5
TECHNICAL SUPPORT AND ORDERING INFORMATION
For technical questions and more detailed information on VWR’s products please visit www.de.vwr.com
to find the respective contact person.
Power Supplies
Do not hesitate to contact us for advice on which Power Supply is most suitable for your application.
PerfectBlue™ Double Gel System Twin S
Item Description Cat. No.
Double gel system Twin S system for gels 10 x 10 cm (W x L), without casting base PEQL45-1010-I
system for gels 10 x 10 cm (W x L), with casting base PEQL45-1010-C
Casting base S including silicone gaskets and cams PEQL45-1010-CB
Cast-It™ S gel caster complete system for sealing up to 2 gels PEQL45-1010-CU
Silicone gaskets 2 replacement silicone gaskets for Casting base S PEQL45-1010-CG
Replacement gaskets 2 replacement gaskets for upper buffer chamber S PEQL45-1010-GK
Side clamps 2 side clamps for fixation of glass plates PEQL45-1010-CL
Wing screws 2 wing screws for fixation of side clamps PEQL45-1010-WK
Notched glass plate clear, 10 x 10 x 0.24 cm PEQL45-1010-10R
Blank glass plate clear, 10 x 10 x 0.24 cm PEQL45-1010-10G
Notched alumina plate white, 10 x 10 x 0.10 cm PEQL45-1010-10RA
Standard combs 1.5 mm 6 teeth 175 µl* PEQL45-1010-6D
1.5 mm 8 teeth 120 µl* PEQL45-1010-8D
1.5 mm 10 teeth 90 µl* PEQL45-1010-10D
1.5 mm 12 teeth 70 µl* PEQL45-1010-12D
0.8 mm 6 teeth 95 µl* PEQL45-1010-6C
0.8 mm 8 teeth 65 µl* PEQL45-1010-8C
0.8 mm 10 teeth 50 µl* PEQL45-1010-10C
0.8 mm 12 teeth 35 µl* PEQL45-1010-12C
Preparative combs 1.5 mm 2 teeth 1490/105 µl* PEQL45-1010-PD
0.8 mm 2 teeth 795/56 µl* PEQL45-1010-PC
SpacerPlacer Facilitates the correct alignment of Spacers PEQL45-1010-SP
Spacer Sets
(2 side & 1 bottom spacer)
1.5 mm 0.7 cm wide 3 pieces PEQL45-1010-SD
0.8 mm 0.7 cm wide 3 pieces PEQL45-1010-SC
* volumes are calculated for a well length of 14 mm

Instruction Manual Cast-It™ Gel CASTER
VWR v0617 E 6
PerfectBlue™ Double Gel System Twin M
Item Description Cat. No.
Double gel system Twin M system for gels 16 x 14 cm (W x L), without casting base PEQL45-1614-I
system for gels 16 x 14 cm (W x L), with casting base PEQL45-1614-C
Casting base M including silicone gaskets and cams PEQL45-1614-CB
Cast-It™ M gel caster complete system for sealing up to 2 gels PEQL45-1614-CU
Silicone gaskets 2 replacement silicone gaskets for Casting base M PEQL45-1614-CG
Replacement gaskets 2 replacement gaskets for upper buffer chamber M PEQL45-1614-GK
Side clamps 2 side clamps for fixation of glass plates PEQL45-1614-CL
Wing screws 2 wing screws for fixation of side clamps PEQL45-1010-WK
Notched glass plate clear, 16 x 14 x 0.32 cm PEQL45-1614-14R
Blank glass plate clear, 16 x 14 x 0.32 cm PEQL45-1614-14G
Standard combs 1.5 mm 10 teeth 234 µl* PEQL45-1614-10D
1.5 mm 15 teeth 140 µl* PEQL45-1614-15D
1.5 mm 20 teeth 90 µl* PEQL45-1614-20D
1.5 mm 24 teeth 65 µl* PEQL45-1614-24D
0.8 mm 10 teeth 125 µl* PEQL45-1614-10C
0.8 mm 15 teeth 75 µl* PEQL45-1614-15C
0.8 mm 20 teeth 50 µl* PEQL45-1614-20C
0.8 mm 24 teeth 35 µl* PEQL45-1614-24C
Preparative combs 1.5 mm 2 teeth 3570/150 µl* PEQL45-1614-PD
0.8 mm 2 teeth 1900/80 µl* PEQL45-1614-PC
SpacerPlacer Facilitates the correct alignment of Spacers PEQL45-1010-SP
Spacer Sets
(2 side & 1 bottom spacer)
1.5 mm 1.0 cm wide 3 pieces PEQL45-1614-SD
0.8 mm 1.0 cm wide 3 pieces PEQL45-1614-SC
* volumes are calculated for a well length of 20 mm
PerfectBlue™ Double Gel System Twin L
Item Description Cat. No.
Double gel system Twin L system for gels 20 x 20 cm (W x L), without casting base PEQL45-2020-I
system for gels 20 x 20 cm (W x L), with casting base PEQL45-2020-C
Casting base L including silicone gaskets and cams PEQL45-2020-CB
Cast-It™ L gel caster complete system for sealing up to 2 gels PEQL45-2020-CU
Silicone gaskets 2 replacement silicone gaskets for Casting base L PEQL45-2020-CG
Replacement gaskets 2 replacement gaskets for upper buffer chamber L PEQL45-2020-GK
Side clamps 2 side clamps for fixation of glass plates PEQL45-2020-CL
Wing screws 2 wing screws for fixation of side clamps PEQL45-1010-WK
Notched glass plate clear, 20 x 20 x 0.32 cm PEQL45-2020-20R
Blank glass plate clear, 20 x 20 x 0.32 cm PEQL45-2020-20G
Standard combs 1.5 mm 10 teeth 305 µl* PEQL45-2020-10D
1.5 mm 15 teeth 185 µl* PEQL45-2020-15D
1.5 mm 20 teeth 125 µl* PEQL45-2020-20D
1.5 mm 25 teeth 90 µl* PEQL45-2020-25D
0.8 mm 10 teeth 165 µl* PEQL45-2020-10C
0.8 mm 15 teeth 100 µl* PEQL45-2020-15C
0.8 mm 20 teeth 65 µl* PEQL45-2020-20C
0.8 mm 25 teeth 50 µl* PEQL45-2020-25C
Preparative combs 1.5 mm 2 teeth 3330/110 µl* PEQL45-2020-PD
0.8 mm 2 teeth 1776/59 µl* PEQL45-2020-PC
SpacerPlacer Facilitates the correct alignment of Spacers PEQL45-1010-SP
Spacer Sets
(2 side & 1 bottom spacer)
1.5 mm 1.0 cm wide 3 pieces PEQL45-2020-SD
0.8 mm 1.0 cm wide 3 pieces PEQL45-2020-SC
* volumes are calculated for a well length of 20 mm

Instruction Manual Cast-It™ Gel CASTER
VWR v0617 E 7
PerfectBlue™ Double Gel System Twin ExW S
Item Description Cat. No.
Double gel system
Twin ExW S
system for gels 20 x 10 cm (W x L), without casting base PEQL45-2010-I
system for gels 20 x 10 cm (W x L), with casting base PEQL45-2010-C
Casting base ExW including silicone gaskets and cams PEQL45-2010-CB
Cast-It™ ExW S gel caster complete system for sealing up to 2 gels PEQL45-2010-CU
Silicone gaskets 2 replacement silicone gaskets for Casting base ExW S PEQL45-2010-CG
Replacement gaskets 2 replacement gaskets for upper buffer chamber ExW S PEQL45-2010-GK
Side clamps 2 side clamps for fixation of glass plates PEQL45-2010-CL
Wing screws 2 wing screws for fixation of side clamps PEQL45-1010-WK
Notched glass plate clear, 20 x 10 x 0.30 cm PEQL45-2010-10R
Blank glass plate clear, 20 x 10 x 0.30 cm PEQL45-2010-10G
Standard combs 1.5 mm 10 teeth 305 µl* PEQL45-2010-10D
1.5 mm 15 teeth 185 µl* PEQL45-2010-15D
1.5 mm 20 teeth 125 µl* PEQL45-2010-20D
1.5 mm 25 teeth 90 µl* PEQL45-2010-25D
0.8 mm 10 teeth 165 µl* PEQL45-2010-10C
0.8 mm 15 teeth 100 µl* PEQL45-2010-15C
0.8 mm 20 teeth 65 µl* PEQL45-2010-20C
0.8 mm 25 teeth 50 µl* PEQL45-2010-25C
Micro titer combs 1.5 mm 18 teeth 105 µl* PEQL45-2010-MTD
1.5 mm 36 teeth 50 µl* PEQL45-2010-MTC
0.8 mm 18 teeth 55 µl* PEQL45-2010-MT2D
0.8 mm 36 teeth 25 µl* PEQL45-2010-MT2C
Preparative combs 1.5 mm 2 teeth 3330/110 µl* PEQL45-2010-PD
0.8 mm 2 teeth 1776/59 µl* PEQL45-2010-PC
SpacerPlacer Facilitates the correct alignment of Spacers PEQL45-1010-SP
Spacer Sets
(2 side & 1 bottom spacer)
1.5 mm 1.2 cm wide 3 pieces PEQL45-2010-SD
0.8 mm 1.2 cm wide 3 pieces PEQL45-2010-SC
* volumes are calculated for a well length of 14 mm