Bio-Rad Criterion Blotter User Manual
Criterion
™
Blotter
Instruction Manual
Catalog Numbers
170-4070
170-4071
For Technical Service Call Your Local Bio-Rad Office or in the U.S. Call 1-800-4BIORAD (1-800-424-6723)
Table of Contents
Page
Section 1 General Information....................................................................................1
1.1 Introduction ................................................................................................................1
1.2 Specifications .............................................................................................................2
1.3 Safety ..........................................................................................................................2
Section 2 Set Up and Basic Operation........................................................................3
2.1 Preparation for Blotting..............................................................................................3
2.2 Begin Transfer............................................................................................................5
2.3 Acidic Transfers .........................................................................................................6
Section 3 Transfer Conditions .....................................................................................6
3.1 General Guidelines to Transfer Buffers and Running Conditions ............................6
3.2 Notes on Electrophoretic Transfer Conditions ..........................................................8
Section 4 Strategies For Optimizing Electro-Elution .............................................11
4.1 Optimizing Protein Transfer ....................................................................................11
4.2 Optimizing DNA and RNA Transfer.......................................................................12
Section 5 Choice of Blotting Membranes.................................................................12
5.1 Protein Blotting Membranes ....................................................................................12
5.2 DNA and RNA Blotting Membranes ......................................................................13
Section 6 Troubleshooting Guide ..............................................................................14
6.1 Electrophoretic Transfer...........................................................................................14
Section 7 Maintenance ...............................................................................................16
Section 8 Product Information..................................................................................16
Section 9 References ...................................................................................................17
Section 10 Warranty.....................................................................................................19
Section 1
General information
1.1 Introduction
The Criterion Blotter is an electrophoretic transfer cell designed for use with Criterion
precast gels as well as hand cast gels. The Criterion Blotter cell is available with standard
wire platinum electrodes or with plate electrodes, which consist of a platinum-coated titanium anode and a stainless steel cathose. Transfers are performed with either set of electrodes
positioned 4.3 cm apart, using one or two gel holder cassettes positioned between the electrodes. This allows generation of ahigh intensity electrical field for and efficient transfer when
used in combination with the PowerPac 200 Power Supply. Cooling is required for temperature control and can be achieved with the sealed ice block included with the cell, or with the
optional cooling coil. The Criterion Gel Blot Assembly Tray provides for lab tidy assembly
of gel blot sandwiches and gel soaking, while minimizing the possibility of incorrect sandwich
assembly. The foller is useful to ensure proper contact and removal of trapped bubbles during sandwich assembly.
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1.2 Specifications
Criterion Blotter tank
Overall dimensions 11.8 x 21.8 x 15 cm
Material Molded polysulfone
Buffer requirement 1.3 liters
Electrodes
Electrode Dimension 9.45 x 13.84 cm
Material Support card Red or black molded polysulfone
Anode plate Platinum coated titanium
Cathode plate Stainless steel
Wire electrodes Platinum wire
Distance Anode to Cathode 4.3 cm
Cassettes
Cassette dimension 11.4 x 16.5 cm
Material Red or black molded polysulfone
Maximum gel size 9.4 x 15 cm
Gel Capacity 2 Criterion or 4 Ready Gel precast gels
Gel/Blot Assembly tray
Material Molded polycarbonate
Overall dimensions 17.3 x 32.5 x 5.7 cm
Assembly compartment 14.3 x 17.3 x 3.2 cm
Soaking compartment 12.1 x 17.3 x 3.2 cm
1.3 Safety
The instrument is for research purposes only.
Power to the Criterion Blotter cell is supplied by an external DC voltage power supply.
This power supply must be ground isolated in such a way that the DC voltage output floats
with respect to ground. All of Bio-Rad’s power supplies meet this important safety
requirement. Regardless of which power supply is used, the maximum specified operating
parameters for the cell are:
300 VDC Maximum voltage limit
200 Watts Maximum power limit
50 °C Maximum ambient temperature limit
Current to the cell, provided from the external power supply, enters the unit through the
lid assembly, providing a safety interlock to the user. Current to the cell is broken when the
lid is removed. Do not attempt to circumvent this safety interlock, and always turn the power
supply off before removing the lid, or when working with the cell in any way.
The Criterion Blotter is certified to meet EN61010-1* safety standard for safety of
laboratory equipment. Certified products are safe to use when operated in accordance with the
instruction manual. This safety certification does not extend to other equipment or
accessories not EN61010-1 certified, even when connected to the Criterion Blotter.
This instrument should not be modified or altered in any way. Alteration of this instrument
will void the manufacturer's warranty, void the EN61010-1 safety certification and create a
potential safety hazard for the user.
Bio-Rad is not responsible for any injury or damage caused by the use of this instrument
for purposes other than for which it is intended or by modifications of the instrument not
performed by Bio-Rad or an authorized agent.
Section 2
Set Up and Basic Operation
Criterion Blotter Cell Assembly
2.1 Preparation for Blotting
1. Freeze ice block prior to preparation of blot assembly.
2. Prepare the transfer buffer. (See Section 3.3 for buffer formulation. Using buffer
pre-chilled to 4°C will improve heat dissipation.)
3. Cut membrane and the filter paper to the dimensions of the gel if necessary.
4. Equilibrate gel in transfer buffer for 15 minutes.
Note: Always wear clean gloves when handling the transfer cell, membranes, filter paper, or
gels to prevent contamination. (See Sectioin 3.2 for details.)
5. Set up transfer apparatus.
• Fill the Criterion Blotter tank with transfer buffer to about 50% of the fill volume.
• Place a magnetic stir bar inside the tank. (0.8–10 mm)
• Place the ice block in the ice block pocket in the back of the cell. Flip down the lever
to hold the ice block down.
• Alternatively, the optional cooling coil can be used. Connect it to an appropriate
recirculating water chiller and place it in the grooves in the back of the tank.
New Figure.
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4. Set up the gel/membrane sandwich:
a. Pour chilled transfer buffer into each compartment of the gel/blot assembly tray.
b. Place the membrane (nitrocellulose, PVDF, etc.) in the front/small compartment of
the tray. Let it soak while you set up steps c–g.
c. Place the cassette in the back/large compartment of the tray: Open the cassette so
that the red side with the handle is vertical (anode) and the black side (cathode) is
laying horizontal and submerged in transfer buffer.
d. Place a fiber pad on top of the black side of the cassette, submerged in buffer. Push
on the fiber pad with gloved finger tips to thoroughly soak the pad.
e. Place a piece of filter paper on top of the fiber pad (it will wet immediately).
f. Gently place the pre-equilibrated gel on top of the filter paper. Use the roller to remove
any air bubbles that may be trapped underneath the gel.
g. Take the membrane from the front compartment and place it on top of the gel taking
care not to trap any air bubbles. The membrane should not be moved or adjusted
once it touches the gel because this can cause data ghost prints and artifacts. If you
feel that you must adjust the membrane placement, use a fresh pre-wetted membrane.
Use the roller to roll out bubbles (see figure).
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Fiber pad
Blot absorbant paper
Membrane
Gel
Blot absorbant paper
Fiber pad
h. Place a piece of filter paper on top of the membrane. Run the roller gently over the
top of the filter paper to remove any air bubbles trapped in the sandwich.
i. Wet a second fiber pad in the front compartment of the tray (where the membrane was
soaking) again using finger tips to completely saturate the pad with transfer buffer.
Then place the wet fiber pad on top of the second filter paper.
j. Lower the clamp-side of the cassette, and lock in the closed position (see figure).
Note: New fiber pads will create a tight fit when the cassette is closed. Subsequent use will
make the pads more compressible.
2.2 Begin Transfer
a. Move the locked cassette into the groove in the blotter tank, aligning the red side of
the card with the red electrode. Make sure the magnetic stirrer is free to move (see
figure).
b. After both cassettes are in place, add the remaining transfer buffer to the fill level
marked on the tank.
c. Put on the lid, plug the cables into the power supply, and run the blot. Refer to Section
3 for run times with various buffers.
d. Transfer at a constant voltage using the Bio-Rad PowerPac 200 which has current
capacity of 2A.
Upon completion of the run, disassemble the blotting sandwich and remove the
membrane for development. Clean the cell, fiber pads, and cassettes with multiple
rinses of deionized water.
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