VWR ShiroGel 700-0250 User manual

VWR ShiroGel Horizontal Manual, Ver.1, 08.04.2010
VWR ShiroGel
Horizontal Electrophoresis Gel Boxes
INSTRUCTION MANUAL
European Catalogue Number(s):
7x10cm 700-0250
15x10cm 700-0253
15x15cm 700-0255
Version: 1
Issued: 08.04.2010

VWR ShiroGel Horizontal Manual, Ver.1, 08.04.2010
Legal Address of Manufacturer
United Stat
es
Europe
VWR International VWR International bvba
1310 Goshen Parkway
Researchpark Haasrode 2020
West Chester, PA 19380 Geldenaaksebaan 464
800
-
932
-
5000
B
-
3001 Leuven
http://www.vwr.com + 32 16 385011
http://be.vwr.com
Country of origin
UK

VWR ShiroGel Horizontal Manual, Ver.1, 08.04.2010
Table of Contents
Page
Warning 4
Safety information 4
Package Contents 5
Unpacking 5
Installation 6
Intended Use 7
Symbols and conventions 7
Description of Buttons and Switches
7
Instructions for use 8
Troubleshooting 10
General maintenance 11
Technical Service 12
Warranty 12
Disposal 13
Appendix 14
Accessories 15

VWR ShiroGel Horizontal Manual, Ver.1, 08.04.2010
Warning
When used correctly, these units pose no health risk. However, these units can deliver dangerous
levels of electricity and are to be operated only by qualified personnel following the guidelines laid
out in this instruction manual.
Anyone intending to use this equipment should read the complete manual thoroughly.
Safety Information
To avoid electrical shock:
The unit must never be used without the safety lid correctly in position.
To avoid electrical shock:
The unit should not be used if there is any sign of damage to the external tank or lid.
These units comply with the statutory CE safety directives:
Low Voltage Directive: 2006/95/EC RoHS Directive: 2002/95/EC
EMC Directive: 2004/108/EC WEEE 2006/96/EC
EN 61010-1:1993/BS EN 61010-1:1993
Avoiding Damage to the Instrument
The units should never come into contact with the following cleaning agents, these will
cause irreversible and cumulative damage:-
Acetone, Phenol, Chloroform, Carbon tetrachloride, Methanol, Ethanol, Isopropyl alcohol Alkalis.
Rnase Decontamination
This can be performed using the following protocol:-
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 wear gloves and safety glasses.
RNaseZAP™ (Ambion) can also be used. Please consult the instructions for use with acrylic gel
tanks.

VWR ShiroGel Horizontal Manual, Ver.1, 08.04.2010
Package Contents
VWR ShiroGel 7x10cm
Units include tank, lid and electrodes and include the following accessories:-
Buffer Volume 225 ml
Tray Tray
Dams
Combs Loading Guides Cables
VWR
ShiroGel
7x10cm
7 x 10cm
(W x L) Pack of 2 3 x 1.5mm, 8 sample
Well Visualization
Strips
Black
Red
VWR ShiroGel 15x10cm and 15x15cm
Units include tank, lid and electrodes and include the following accessories:-
Buffer volume 500 ml
Tray Tray Dams Combs Loading Guides Cables
VWR
ShiroGel
15x10cm 15 x 10cm
(W x L) Pack of 2
1 x 1.5mm, 20
sample, 1 x
1.5mm, 16
sample
(Multichannel)
Well Visualization
Strips
Black
Red
VWR
ShiroGel
15x15cm
15 x 15cm
(W x L) Pack of 2
1 x 1.5mm, 20
sample, 1 x
1.5mm, 16
sample
(Multichannel), I x
1.5mm, 28
sample
(Multichannel)
Well Visualization
Strips
Black
Red
Unpacking
These product descriptions and included component information should be referred to as soon as
the units are received to ensure that all components have been included. The unit should be
checked for damage when received. Please contact your supplier if there are any problems or
missing items.

VWR ShiroGel Horizontal Manual, Ver.1, 08.04.2010
Installation
Usage Guidance and restrictions:
• Maximum altitude 2,000m.
• Temperature range between 4°C and 60°C.
• Maximum relative humidity 80% for temperatures up to 31°C decreasing linearly to 50% relative
humidity at 40°C.
• Not for outdoor use.
This apparatus is rated POLLUTION DEGREE 2 in accordance with IEC 664. POLLUTION
DEGREE 2, states that: “Normally only non-conductive pollution occurs. Occasionally, however,
a temporary conductivity caused by condensation must be expected.”
Setting up the ShiroGel Gel Tanks:-
Instructions for attaching Electrode Leads
1. Note the position of the lid on the unit. This shows the correct polarity and the correct
orientation of the cables, black is negative and red positive.
2. Remove the lid from the unit. Note if the lid is not removed, fitting the cables may result in
un-tightening of the gold plug and damage to the electrode.
3. Screw the cables into the tapped holes as fully as possible so that there is no gap
between the lid and the leading edge of the cable fitting.
4. Refit the lid.
Instructions for using Gel Visualization Strips
These can be fitted to enhance visibility of the wells if desired. They can be fitted to the
unit.
1. Seat the tray in the unit and note the position of the comb grooves. The samples run
black to red but the trays can be used frontward or backwards so ensure that the comb
grooves closest to the black electrode are marked.
2. Remove the tray.
3. Peel the back off of the loading guide and carefully apply the loading guide directly to the
gel box.
The unit is now ready for use.

VWR ShiroGel Horizontal Manual, Ver.1, 08.04.2010
Intended use
For research use only. Not intended for any animal or human therapeutic or diagnostic use.
Symbols and conventions
The following chart is an illustrated glossary of the symbols that are used in this manual.
CAUTION This symbol indicates a potential risk and alerts you to
proceed with caution
CAUTION This symbol indicates the presence of high voltage and
warns the user to proceed with caution
Description of Buttons and Switches
Replaceable electrodes
Electrode wires
Height adjustable combs
Rubber casting dams

VWR ShiroGel Horizontal Manual, Ver.1, 08.04.2010
Getting Started
Gel Preparation:-
1. Table 1 shows the volume of agarose solution required to make the desired agarose gel for
each unit tray size. For a standard 0.7% agarose gel, add 0.7 grams of agarose to 100 ml of 1x
TAE or TBE solution. The same 1 x solution should be used in the tank buffer solution.
Table 1.
VWR ShiroGel 7x10cm VWR ShiroGel 15x10cm VWR ShiroGel 15x15cm
Tray Gel volume for
a 1mm thick
gel
Tray Gel volume for
a 1mm thick
gel
Tray Gel volume for
a 1mm thick
gel
7 x10cm 70ml 15 x 10cm 150ml 15 x 15cm 225ml
2. Add the agarose powder to a conical flask.
3. Add the appropriate amount of 1x TAE or TBE solution from the table above. To prevent
evaporation during the dissolving steps below, the conical flask should be covered.
4. Dissolve the agarose powder by heating the agarose either on a magnetic hot plate with
stirring bar or in a microwave oven. If using the microwave method, mark the level of the solution
on the container. The microwave should be set at around a 400 watt or medium setting and the
flask swirled every minute. The solution should be heated until all crystals are dissolved. This is
best viewed against a light background. Crystals appear as translucent crystals. These will
interfere with sample migration if not completely dissolved. Once the once the agarose is
completely dissolves add ddi water to the solution to the marked level.
The gel must be cooled to between 50°C and 60°C degrees before pouring.
Gel Pouring:-
The VWR systems allow two different methods of gel casting:-
Casting Dams and Traditional Tape
Using trays with Casting Dams
1. Fit the casting dams over each end of the tray and place onto a level surface. The dams
should be fitted so that there is no gap between the sides of the tray and the groove in
the dams. This will ensure that there is no possibility of gel leakage.

VWR ShiroGel Horizontal Manual, Ver.1, 08.04.2010
2. Place the comb(s) in the grooves. Each tray has more than one comb grove so that
multiple combs can be used. Using multiple combs increases sample number available
per gel but decreases run length and care must be taken to ensure that samples from the
first wells do not migrate into the lanes of the second comb wells.
3. Pour in the agarose carefully so as not to generate bubbles. Any bubbles that do occur
can be smoothed to the edge of the gel and dispersed using a pipette tip.
4. Allow the agarose to set, on a flat surface; the agarose will become opaque once
solidified.
5. Carefully remove the gel casting gates and comb and transfer the gel including tray to the
main tank.
Using Traditional tape method:-
1. Autoclave or plastic backed general tape should be used. A length 5cm longer than the
width of each end of the tray should be cut. One length should be placed over one end of
the tray and stuck m1cm in from the tray edge. This should then be folded and the edges
sealed securely. Repeat for the other end and place onto a level surface for gel pouring.
2. Place the comb(s) in the grooves. Each tray has more than one comb grove so that
multiple combs can be used. Using multiple combs increases sample number available
per gel but decreases run length and care must be taken to ensure that samples from the
first wells do not migrate into the lanes of the second comb wells.
3. Pour in the agarose carefully so as not to generate bubbles. Any bubbles that do occur
can be smoothed to the edge of the gel and dispersed using a pipette tip.
4. Allow the agarose to set, on a flat surface; the agarose will become opaque once
solidified.
5. Carefully remove the gel casting gates and comb and transfer the gel including tray to the
main tank.
Running the Gel:-
1. Mix the sample to be loaded with sample buffer – see page 9 solutions for common sample
buffers. Usually 3ul of sample buffer is adequate but less may be used with sample volumes of
less than 10ul.
2. Fill the unit with buffer until the gel is just flooded with buffer. This will give the fastest
resolution times. For enhanced quality of resolution of sample, fill the unit to 5mm above the gel.
Note: DNA, RNA have a net negative charge and will migrate towards the anode.
3. Load the samples into the wells using pipettes. Multi-channel pipettes can be used for
loading samples with MC compatible combs, see listing in accessories for identification of these.

VWR ShiroGel Horizontal Manual, Ver.1, 08.04.2010
4. Carefully place the lid on the tank and connect to a power supply.
5. Typically gels are run at between 90 and 150 volts. However, maximum voltages are indicated
on the serial badge of each unit. It should be noted that higher voltages generally give faster but
poorer quality sample resolution.
NOTE: Use only power supplies equipped with no-load and ground fault detection
Gel Staining and Viewing:-
1. Transfer the gel to a vessel containing the appropriate volume of 0.5 µg/ml ethidium bromide
stain for 15–30 minutes, see solutions for stock stain concentration and adjust to the volume used
accordingly. The entire gel should be covered.
NOTE: Ethidium bromide is a suspected carcinogen and the necessary safety precautions
should be undertaken. Always wear protective clothing, gloves and safety glasses
2. De-stain the gel for 10–30 minutes in distilled water again ensuring the gel is completely
immersed.
3. Rinse the gel twice for a couple of seconds with distilled water.
4. Transfer the gel to a UV Transilluminator.
5. The samples will often appear as brighter, clearer bands when photographed or viewed using a
gel documentation system. However if the gel bands are too faint then the staining procedure
should be adjusted so that there is less de-staining. If there is too much background then the
staining procedure should be adjusted so that there is more de-staining.