Bio-Rad ReadySub-Cell GT Cells User Manual
Sub-Cell® GT Agarose Gel
Electrophoresis Systems
Instruction Manual
Catalog # 170-4401 to 170-4406
170-4481 to 170-4486
Warranty
Bio-Rad Laboratories warrants the Sub-Cell GT, Wide Mini-Sub® Cell GT, and Mini-Sub Cell GT
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
To insure the best performance from the Sub-Cell GT 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 GT 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. _____________________________________
For any inquiry or request for repair service, contact Bio-Rad Laboratories after confirming the model
and serial number of your instrument.
Table of Contents
Page
Section 1 General Information 1
1.1 Introduction 1
1.2 Safety 1
1.3 System Components 2
1.4 Specifications 4
Section 2 Operating Instructions 4
2.1 DNA Gel Preparation 4
2.2 Casting Agarose Gel Slabs 6
2.3 Electrophoresis 8
2.4 Nucleic Acid Staining and Visualization 9
2.5 Note on Blotting 10
Section 3 Gel and Electrophoresis Reagent Preparation 10
Section 4 Care and Maintenance 11
4.1 Cleaning Sub-Cell GT Components 11
4.2 Compatible Cleaning Agents 11
4.3 Maintenance Schedule 12
4.4 Electrode Replacement 12
4.5 RNase Decontamination 13
Section 5 Troubleshooting 13
Section 6 Product Information 14
6.1 Sub-Cell GT Systems 14
6.2 Sub-Cell GT System Accessories 15
6.3 Related Bio-Rad Products 17
Section 7 References 19
Section 1
!
!
General Information
1.1 Introduction
The Sub-Cell® GT instruments (basic Sub-Cell® GT cell, Wide Mini-Sub® Cell GT, and Mini-Sub Cell®
GT) comprise a comprehensive and flexible 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 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 gel submarine electrophoresis can effectively separate nucleic acids from
20 base pairs to 20 kilobase pairs in length.
The Sub-Cell GT systems are designed for years of reproducible and 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. Gels can also be cast in the GT bases using specially
designed casting gates. Replaceable electrode cassettes provide a simple way to replace electrode wires.
A comprehensive assortment of base and tray sizes, including a variety of preparative, analytical, and
multichannel pipet compatible combs, makes these systems ideal for any agarose gel application.
Note: This manual contains instructions for the Sub-Cell GT electrophoresis systems only. Prior to the
release of the Sub-Cell GT systems, Bio-Rad supplied similar agarose gel electrophoresis cells: the original
Sub-Cell DNA electrophoresis cell, Wide Mini-Sub cell, and Mini-Sub cell systems. This manual does not
provide information on these earlier versions. Contact your local Bio-Rad representative for information
concerning the original Sub-Cell systems.
Definition of Symbols
Caution, risk of electrical shock Caution (refer to accompanying documents)
1.2 Safety
The Sub-Cell GT electrophoresis systems are designed for maximum user safety. The buffer chambers
are made of 2.75 mm thick injection-molded acrylic to create a leak-free electrophoresis environment.
The safety lids surround the buffer chamber to protect the user from exposure to electrical currents.
All Sub-Cell GT systems were designed for indoor use only.
Before use, inspect the GT base for cracks or chips, which may allow the buffer to leak from the base
and cause a potential electrical hazard. Additionally, inspect all electrical cables, banana jacks, 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 hazard. 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 local Bio-Rad representative.
1
Power to Sub-Cell GT units 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. The recommended power supply for these units is
the PowerPac
no load, overload, rapid resistance change, and ground leak detection capabilities. The maximum specified
operating parameters for the Sub-Cell GT systems are given in Table 1.1.
Table 1.1 Sub-Cell GT systems operating parameters
Sub-Cell Wide Mini-Sub Mini-Sub
GT Cell Cell GT Cell GT
Maximum voltage limit 200 VDC 150 VDC 150 VDC
Maximum power limit 40 Watts 45 Watts 10 Watts
Maximum Buffer temperature 40°C 40°C 40°C
which provides a safety interlock. 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.
certified products are safe to use when operated in accordance with the instruction manual. This instrument
should not be modified in any way. Alteration of this instrument will:
• Voidthemanufacturer’swarranty
• VoidtheIEC1010-1safetycertification
• Createapotentialsafetyhazard
of 4°C and 40°C and altitudes up to 2,000 meters; safe at a maximum relative humidity of 80% for
temperatures up to 31°C, and for temperatures higher than 31°C, safe at a maximum humidity that
decreases linearly to 50% at 40°C. Bio-Rad is not responsible for any injury or damage caused by the use of
this instrument for purposes other than those for which it is intended, or by modifications of the instrument
not performed by Bio-Rad or an authorized agent. There are no user-serviceable parts in this apparatus.
To insure electrical safety, do not attempt to service this apparatus.
™
Basic power supply. The PowerPac™ Basic power supply contains safety features such as
Electric current to the cell, from the external power supply, enters the unit through the lid assembly,
Important: These Bio-Rad instruments are certified to meet IEC 1010-1** safety standards. IEC-
The Sub-Cell GT systems conform to IEC-1010-1 standards: safe between the operating temperatures
1.3 System Components
Each of the Sub-Cell GT systems comes with the components listed in Table 1.2 (see Figure 1.1, page
3, for part description). Check your instrument to be sure 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.
Table 1.2 Sub-Cell GT system components
Sub-Cell GT Wide Mini-Sub Cell Mini-Sub Cell
System GT System GT System
Item Quantity Quantity Quantity
GT Base (buffer chamber) 1 1 1
Gel Casting Gates (optional) 2 2 2
Safety Lid and Cables 1 1 1
UVTP Gel Tray 1 1 1
Fixed Position Comb 2 2 2
(15 well, 1.5 mm thick) (15 well, 1.5 mm thick) (8 well, 1.5 mm thick)
(20 well, 1.5 mm thick) (20 well, 1.5 mm thick) (15 well, 1.5 mm thick)
Leveling Bubble 1 1 1
Gel Caster (optional) 1 1 1
Instruction Manual 1 1 1
2
Electrical cables
Safety lid
Electrical leads
Comb
slots
Banana plug/
electrode
wire assembly
Fixed height
comb
Fixed height
comb
Fluorescent
ruler
UV-transparent
gel tray
Gel casting gates
Mini-Sub Cell GT Base
Safety lid removal tab
Gel stage
Fluorescent
ruler
Cam lever
Leveling bubble
Leveling feet
Fig.1.1. Mini-Sub Cell GT components.
UV-transparent gel tray
Gel caster
3
1.4 Specifications
Sub-Cell GT Wide Mini-Sub Mini-Sub Cell
System Cell GT System GT System
GT base footprint (L x W x H) 40.5 x 18 x 9.4 cm 25.5 x 17.8 x 6.8 cm 25.5 x 9.2 x 5.6 cm
GT base buffer volume
GT base gel size 15 x 15 cm 15 x 7 cm 7 x 7 cm
Gel tray sizes 15 x 10 cm 15 x 7 cm 7 x 7 cm
15 x 15 cm 15 x 10 cm 7 x 10 cm
15 x 20 cm
15 x 25 cm
Construction
GT base Molded clear plastic
Gel casting gates Aluminum
Safety lid Molded clear plastic
Electrode wire guard Molded 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 Molded plastic and machined acrylic
Gel casting device Polycarbonate
0.64 cm silicon foam
=
GT base buffer volumes will vary depending on the size and thickness of the gel used.
=
1,500—2,000 ml 650—900 ml 265—320 ml
Section 2
Operating Instructions
Note: See Section 3, Gel and Electrophoresis Reagent Preparation, for information on the 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, page 5, 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) required to make the desired agarose gel concentration
and volume. Use Tables 2.1 and 2.2, page 5, 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 1x electrophoresis buffer.
4
Loading...