Bio-Rad Sub-Cell Model 192 Cell User Manual
Sub-Cell®Model 96 and
Model 192
Agarose Gel
Electrophor esis Systems
Instruction Manual
Catalog Numbers
170-4500 through 170-4511
For Technical Service Call Your Local Bio-Rad Office or in the U.S. Call 1-800-4BIORAD (1-800-424-6723)
Warranty
Bio-Rad Laboratories warrants the Sub-Cell Model 96 and Model 192 electrophoresis systems against
defects in materials and workmanship for 1 year. If any defects occur in the instrument during this
warranty period, Bio-Rad Laboratories will repair or replace the defective parts free. The following
defects, however, are specifically excluded:
1. Defects caused by improper operation.
2. Repair or modification done by anyone other than Bio-Rad Laboratories or an authorized
agent.
3. Use of fittings or other spare parts supplied by anyone other than Bio-Rad Laboratories.
4. Damage caused by accident or misuse.
5. Damage caused by disaster.
6. Corrosion due to use of improper solvent or sample.
This warranty does not apply to parts listed below:
1. Platinum Electrode Wires
For any inquiry or request for repair service, contact Bio-Rad Laboratories after confirming the
model and serial number of your instrument.
To insure the best performance from the Sub-Cell electrophoresis systems, become fully acquainted
with these operating instructions before use. Bio-Rad recommends that you first read these instructions
carefully. Assemble and disassemble the unit completely without casting a gel. After these preliminary steps, you should be ready to cast and run a gel.
Bio-Rad also recommends that all Sub-Cell system components and accessories be inspected for
damage, cleaned as recommended in this manual and rinsed thoroughly with distilled water before
use. Record the following for your records:
Model
Catalog No.
Date of Delivery
Warranty Period
Serial No.
Invoice No.
Purchase Order No.
Table of Contents
Page
Warranty Information ..........................................................................Inside Front Cover
Section 1 General Information....................................................................................1
1.1 Introduction ................................................................................................................1
1.2 Safety..........................................................................................................................1
1.3 List of System Parts....................................................................................................3
1.4 Specifications .............................................................................................................5
Section 2 Operating Instructions................................................................................6
2.1 DNA Gel Preparation.................................................................................................6
2.2 Comb Set-up...............................................................................................................7
2.3 Casting Agarose Gels.................................................................................................8
2.4 Recirculation Ports...................................................................................................11
2.5 Electrophoresis.........................................................................................................12
2.6 Nucleic Acid Staining and Visualization.................................................................13
2.7 Note on Blotting.......................................................................................................14
Section 3 Gel and Electrophoresis Reagents Preparation......................................14
3.1 Electrophoresis Buffer Preparation..........................................................................14
3.2 DNA and RNA Gel Preparation ..............................................................................15
3.3 RNA Sample Preparation.........................................................................................15
3.4 DNA and RNA Sample Loading Dye .....................................................................16
3.5 Gel Staining Solution...............................................................................................16
Section 4 Care and Maintenance..............................................................................16
4.1 Cleaning Sub-Cell System Components..................................................................16
4.2 Compatible Cleaning Agents...................................................................................16
4.3 Maintenance Schedule .............................................................................................17
4.4 Electrode Replacement.............................................................................................17
4.5 RNase Decontamination ..........................................................................................18
Section 5 Troubleshooting..........................................................................................19
Section 6 Ordering Information ...............................................................................20
6.1 Sub-Cell Model 96 and 192 Systems.......................................................................20
6.2 Sub-Cell Model 96 and 192 System accessories.....................................................20
6.3 Related Bio-Rad Products........................................................................................21
Section 7 References...................................................................................................25
i
Section 1
General Information
1.1 Introduction
The Sub-Cell instruments comprise a comprehensive and versatile gel electrophoresis
system that effectively separates nucleic acids using submerged agarose gels. Submarine
agarose gels are easy to cast and readily dissipate heat. These gels allow sample underlaying
and also prevent electrical field discontinuities caused by wicks or sample well dehydration.
Agarose gels are ideal for the separation of DNA restriction digestions, Polymerase Chain
Reaction (PCR)* amplified fragments, and genomic DNA and RNA prior to Southern or
Northern blotting. If operated correctly, agarose gels can effectively separate nucleic acids
from 20 base pairs to 20 kilobase pairs in length.
The Sub-Cell Model 96 and 192 electrophoresis cells have been created specifically for
multiple sample analysis. The width of each cell and the analytical combs were designed
based on the fixed spacing of multichannel pipets used to transfer samples from standard
microplates. Forty eight nucleic acid samples (plus three DNA size standards) can be visualized
in one row using the 51-well comb. If two combs are used, samples from all 96 wells of a
microplate can be analyzed on the Model 96. Four or more combs can be used on the
Model 192 for even higher throughput. The Model 96 can run gels 10 or 15 cm in length,
whereas the Model 192 can run gels 10, 15, 20 or 25 cm in length for the analysis of more samples or applications such as genomic DNA separations for Southern blotting.
The Sub-Cell systems give years of rigorous use. These rugged systems incorporate many
features that make casting and running agarose gels simple and efficient. The gel caster provides tape-free gel casting in trays and gels can be cast in the Sub-Cell bases using the gel
casting gates. Replaceable electrode cassettes provide a simple way to replace electrode wires.
A comprehensive assortment of tray sizes and multichannel pipet-compatible combs make
these systems ideal for most high throughput agarose gel applications. Recirculation ports
are provided to prevent heat and pH effects during high voltage or extended run electrophoresis.
Note: This manual contains instructions for the Sub-Cell Model 96 and Model 192
electrophoresis systems only. Bio-Rad supplies similar but smaller agarose gel
electrophoresis cells: the original Sub-Cell, Wide Mini-Sub Cell, and Mini-Sub Cell systems
and the Sub-Cell GT, Wide Mini-Sub Cell GT and Mini-Sub Cell GT systems. This
manual does not provide information concerning these smaller versions. Contact your local
Bio-Rad representative for information concerning the original Sub-Cell and
Sub-Cell GT systems.
* The Polymerase Chain Reaction (PCR) process is covered by patents owned by Hoffmann-LaRoche. Use of the PCR process
requires a license.
1.2 Safety
The Sub-Cell electrophoresis systems are designed for maximum user safety. The buffer
chambers are made of 1/4-inch (.635 cm) thick cast acrylic to create a leak-free
electrophoresis environment. The safety lids surround the entire buffer chamber to protect the
user from exposure to electrical currents. Sub-Cell systems were designed for indoor use only.
1
Before every use, inspect the base for cracks or chips. Cracks or chips may cause the
buffer to leak from the base and cause a potential electrical hazard. Additionally, inspect all
electrical cables, banana jacks, recirculation port fittings, tubing, and plugs for loose
connections, cracks, breaks or corrosion. Do not use any part that is cracked, charred or
corroded. These parts may also cause a potential electrical shock. Contact your local Bio-Rad
representative before using a part that may be considered hazardous.
During electrophoresis inspect the base and workbench for any signs of buffer leakage.
If leaking buffer is detected disconnect the power to the cell immediately and contact your
Bio-Rad representative.
Power to Sub-Cell units is to be 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 Bio-Rad power supplies meet this important safety requirement. The
recommended power supply for this apparatus is the PowerPac 300. The PowerPac 300
contains several safety features such as no load, overload, rapid resistance change, and ground
leak detection capabilities. The maximum specified operating parameters* for the
Sub-Cell Model 96 and Model 192 systems are:
200 VDC Maximum voltage limit
70 Watts Maximum power limit
50 °C Maximum buffer temperature
4 °C – 40 °C Ambient temperature limits
* IEC 1010-1 certification applies to equipment designed to be safe at the operating parameters listed above. Additionally, both
Sub-Cell Model 96 and Model 192 have a maximum operating relative humidity of 80% for temperatures up to 31 °C decreasing
linearly to 50% relative humidity at 40 °C. Certification is valid when systems are operated at altitudes up to 2000 meters.
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.
Important: These Bio-Rad instruments are designed and certified to meet IEC 1010-1*
safety standards. IEC-certified products are safe to use when operated in accordance with
this instruction manual. This instrument should not be modified in any way. Alteration of
this instrument will:
• Void the manufacturer’s warranty
• Void the IEC 1010-1 safety certification
• Create a potential safety hazard
Bio-Rad is not responsible for any injury or damage caused either 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 any authorized agent. No user-serviceable parts are contained in
this apparatus. To ensure electrical safety, do not attempt to service this apparatus.
* IEC 1010-1 is an internationally accepted electrical safety standard for laboratory instruments.
Definition of Symbols
Caution, risk of electrical shock Caution (refer to accompanying documents)
2
1.3 List of System Parts
Each Sub-Cell system comes with the components listed in Table 1.1. Check your
instrument to insure all items are present. Note any damage to the unit which may have
occurred during shipping. Notify Bio-Rad Laboratories if any items are missing or damaged
(see Figure 1.1 for part descriptions, on the following page).
Table 1.1 Sub Cell System Components
Item Quantity
Base (buffer chamber) 1
Gel Casting Gates 2
Safety Lid and Cables 1
UVTP Gel Tray 1
Comb (51 well, 1.5 mm thick) 1
Comb Holder 1
Leveling Bubble 1
Gel Caster (optional) 1
Instruction Manual 1
3
Figure 1.1 Sub-Cell Model 96 and Model 192 Parts
4
Safety Lid
Electrical
Cables
Recirculation
Port Plugs
Comb Holder
Comb
Electrical
Leads
UVTP Gel Tray
Comb Holder Slot
Fluorescent
Ruler
Banana Plug/Electrode
Wire Assembly
Gel Casting
Gates
Base
(Buffer Chamber)
Leveling
Feet
Figure 1.2 Sub-Cell Model 96 and Model 192 Gel Caster Parts
1.4 Specifications
Sub-Cell Model 96
Base Footprint (L x W x H) 29 .5 cm x 29.0 cm x 9.0 cm
Base Buffer Volume* 2.0 L
Base Gel Size 25 x 10 cm
Gel Tray Sizes 25 x 10 cm
25 x 15 cm
Sub-Cell Model 192
Base Footprint (L x W x H) 39.5 cm x 29.0 cm x 9.0 cm
Base Buffer Volume* 3.0 L
Base Gel Size 25 x 15 cm
Gel Tray Sizes 25 x 10 cm
25 x 15 cm
25 x 20 cm
25 x 25 cm
5
Comb and
Comb Holder
UVTP Gel Tray
Gel Caster Leveling
Bubble
Leveling Feet
Movable Wall
Fixed Wall
Contruction
Base Cast Acrylic
Gel Casting Gates Anodized Aluminum
Safety Cover Cast Acrylic
Banana Plug/Electrode Cassette Polycarbonate
Banana Plugs Gold-Plated Brass, 4.4 cm Length
Electrodes Platinum, 0.25 mm Diameter
Electrical Cables Dual, 20 AWG, Tinned Copper Wire Cable
Flame-Retardant Polyurethane Insulation jacket
Electrical Leads Nickel Silver
Gel Tray UV-Transparent Acrylic Plastic (UVTP)
Combs Machined Acrylic
Comb Holder Polycarbonate
Gel Casting Device Polycarbonate
0.64 cm Silicon Foam
* Base buffer volumes will vary depending on the size and thickness of gel used.
Section 2
Operating Instructions
Note: Refer to Section 3 for information on preparation of RNA gels. See References
1 and 2 for more information on DNA and RNA electrophoresis.
2.1 DNA Gel Preparation
DNA agarose gels can be used to separate and visualize DNA of various sizes. Before
casting an agarose gel, consult Table 2.1 to determine the appropriate percent agarose gel to
use based on the size of DNA to be separated.
Procedure
1. Determine the amount of agarose (grams) and volume needed. Use Tables 2.1 and 2.2
as a guide for agarose concentration and gel volume requirements.
Example: For a 1% agarose gel, add 1 gram of agarose to 100 ml of electrophoresis buffer.
Table 2.1 Gel Concentration Required for DNA Separation
1-2
Gel Concentration % DNA Size (Kbp)
0.50 1 – 30
0.75 0.8 – 12
1.00 0.5 – 10
1.25 0.4 – 7
1.50 0.2 – 3
2-5* 0.01 – 0.5
* Sieving agarose such as Bio-Rad AmpliSize®agarose.
6
Loading...