BIO RAD Gel Doc EZ Instrument Manual

Gel Doc™ EZ Imaging System
with Image Lab
Instrument Guide
Version 6.0
Software
Gel Doc™ EZ Imaging System
with Image Lab
Instrument Guide
Version 6.0
Software
The Bio-Rad Technical Support department in the United States is open Monday through Friday, 5:00 AM to 5:00 PM, Pacific time. Go to consult.bio-rad.com for worldwide technical support.
Phone: 1-800-424-6723, option 2
Web: consult.bio-rad.com
Email: Support@Bio-Rad.com (United States/Canada only)
For technical assistance outside the United States and Canada, contact your local technical support office.
Notice
No part of this publication may be reproduced or transmitted in any form or by any means, electronic or mechanical, including photocopy, recording, or any information storage or retrieval system, without permission in writing from Bio-Rad.
Bio-Rad reserves the right to modify its products and services at any time. This user guide is subject to change without notice. Although prepared to ensure accuracy, Bio-Rad assumes no liability for errors or omissions, or for any damage resulting from the application or use of this information.
Credits
1. Image Lab software is based in part on the work of the Qwt project (http://qwt.sf.net).
2. Image Lab software is based in part on the work of the CImg project (http://cimg.sourceforge.net/). See license for details at: http://www.cecill.info/licences/Licence_CeCILL-C_V1-en.html
3. Image Lab software is based in part on the work of the Independent JPEG Group (http://www.ijg.org/).
Coomassie is a trademark of BASF Aktiengesellschaft.
Coomassie Fluor, SYBR, SYPRO are trademarks of Life Technologies Corporation. Bio­Rad Laboratories, Inc. is licensed by Life Technologies Corporation to sell reagents containing SYBR Green I for use in real-time PCR for research purposes only and to sell SYPRO products for research use only under U.S. Patent Number 5,616,502.
Windows is a trademark of Microsoft Corporation. Mac and Mac OS are trademarks of Apple Inc.
GelGreen and GelRed are trademarks of Biotium, Inc. Krypton is a trademark of Thermo Fisher Scientific Inc.
Criterion Stain Free gels are covered by U.S. Pat. No. 7,569,130.
Copyright © 2017 Bio-Rad Laboratories, Inc. All rights reserved.

Table of Contents

Safety and Regulatory Compliance. . . . . . . . . . . . . . . . . . . . . . . . . . . .5
Safety Use Specifications and Compliance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
Alert Icons . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
Cautions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
Warnings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
Instrument Safety Warnings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
Notice . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
Warranty . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
Chapter 1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .11
Gel Doc EZ Imaging System. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
Technical Specifications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
Image Lab Software Capabilities . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
Gel Documentation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
Molecular Weight Assessment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
Quantitation. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
For More Information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
Chapter 2 Setting Up Gel Doc EZ. . . . . . . . . . . . . . . . . . . . . . . . . . . . . .15
Configuring the Imager . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
Initializing an Additional Tray . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22
Recalibrating Flat Field and Dark Image. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23
Chapter 3 Acquiring Images . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .25
Selecting a Sample Tray . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25
Creating a Protocol . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26
Instrument Guide | iii
Table of Contents
Configuring Acquisition Settings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29
Configuring Analysis Settings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32
Configuring Output Settings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32
Running a Default Protocol . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .33
Running a Protocol . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36
Creating a Custom Application . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .38
Editing a Protocol . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39
Appendix A Maintenance and Specifications . . . . . . . . . . . . . . . . . 41
Cleaning the Sample Trays . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41
UV-B Fluorescent Lamp Replacement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41
Error Messages . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .43
Appendix B Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45
Appendix C Accessories . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47
Ordering Information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .47
Appendix D Using Bio-Rad Stain-Free Technology. . . . . . . . . . . . . 49
Stain-Free Workflow . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50
Electrophoresis with Stain-Free Gels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51
Imaging Gels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51
Imaging Blots. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 52
Glossary . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 53
iv | Gel Doc EZ Imaging System with Image Lab Software

Safety and Regulatory Compliance

The Gel Doc™ EZ imaging system is intended for laboratory use only. To help you make informed decisions about safety, we have provided comprehensive operating procedures and safety information in this manual and on labels affixed to the imager. This information will alert you to any potential hazards. It is the user’s responsibility to read and understand the safety information and use it for safe operation of the system.

Safety Use Specifications and Compliance

The Gel Doc EZ imager is designed and certified to meet EN61010, the internationally accepted electrical safety standard, and EN61326 Class A EMC regulations. Certified products are safe to use when operated in accordance with the instruction manual.
This instrument should not be modified or altered in any way. Modification or alteration of this instrument will
Void the manufacturer’s warranty
Void the regulatory certifications
Create a potential safety hazard
Instrument Guide | 5
Safety and Regulatory Compliance
Fig. 1. Gel Doc EZ imager certification label.
For easy customer access, the serial number appears in two locations on your instrument: on the back panel and inside the front door.
6 | Gel Doc EZ Imaging System with Image Lab Software

Alert Icons

Alert icons call attention to caution and warning paragraphs. The icon indicates the
type of hazard addressed.
Table 1. How alert icons are used in this user guide
Icon Explanation
General
Indicates a potential hazard requiring special attention. This icon is used when the hazard or condition is of a general nature.
Electrical hazard
Indicates a potential hazard requiring special attention when you are working with electricity or electrical equipment.
Extreme heat and flammable materials
Indicates a potential hazard requiring special attention when you are working with extreme heat and flammable materials.
Radiation hazard
Indicates a potential hazard requiring special attention when you are working with UV radiation.
Alert Icons

Cautions

A caution alerts you to take or avoid a specific action that could result in loss of data or damage to the instrument. A caution can also indicate that, if the precaution against a potential hazard is not taken, minor or moderate injury might occur.
Example
Caution: With the exception of cleaning or replacing light bulbs, refer all
servicing to qualified Bio-Rad personnel or their agents.
Instrument Guide | 7
Safety and Regulatory Compliance

Warnings

A warning precedes an action that, if not followed correctly, could cause serious injury or death to the operator, serious or total loss of data, or serious damage to the instrument.
Example
WARNING! This instrument must be connected to an appropriate AC voltage
outlet that is properly grounded
.

Instrument Safety Warnings

Before you operate the instrument, carefully read the contents of Table 2.
Table 2. Safety cautions and warnings for the instrument
Icon Meaning
Caution: With the exception of cleaning or replacing light bulbs, refer all
servicing to qualified Bio-Rad personnel or their agents. If you experience technical difficulties with the instrument, contact Bio-Rad to schedule service.
Caution: If the case interlock is defeated, there is a possibility of UV-B
radiation hazard due to UV-B light exposure. Exercise caution when servicing the instrument.
Caution: Disconnect the AC power cord before removing the instrument
cover.
Warning! This instrument must be connected to an appropriate AC voltage
outlet that is properly grounded.
8 | Gel Doc EZ Imaging System with Image Lab Software

Notice

The Gel Doc EZ imaging system is meant for use by specialized personnel who know the health risks associated with reagents normally used in electrophoresis. Bio-Rad Laboratories, Inc. is not responsible for any injury or damage caused by use of this instrument for purposes other than those for which it is intended, or for instrument modifications not performed by Bio-Rad Laboratories, Inc. or an authorized agent.
potential safety hazard for the user.

Warranty

Each Gel Doc EZ imager is protected by a comprehensive instrument warranty agreement. Please read this manual thoroughly so that you fully understand the coverage provided and are aware of your rights and responsibilities. One of the responsibilities of system ownership is regular maintenance. Following the maintenance instructions provided with this manual will help keep your system and peripherals functioning optimally and will protect your investment. Bio-Rad offers a range of comprehensive service agreements that can be tailored to meet your specific needs. Bio-Rad Laboratories is dedicated to your total satisfaction and will be pleased to answer any questions you might have.
Notice
Alteration voids the manufacturer’s warranty and might create a
Instrument Guide | 9
Safety and Regulatory Compliance
10 | Gel Doc EZ Imaging System with Image Lab Software

1 Introduction

The Gel Doc™ EZ imaging system consists of the Gel Doc EZ imager and Image Lab™ software. The imager connects to a separate computer running Image Lab. This combination creates an automated system for imaging and analyzing gels and blots. Data can be viewed, modified, and reported using Image Lab software.
The Gel Doc EZ imaging system supports multiple applications, including Coomassie and ethidium bromide staining, blue excitation for nondestructive DNA visualization, and stain-free gel and blot imaging.
Image Lab protocols automate sample image acquisition, analysis, and report generation with the push of a button on the imager.
For complete information about Image Lab software, see the Image Lab Software User Guide.

Gel Doc EZ Imaging System

The Gel Doc EZ imager combines a low-noise detection camera with a UV transilluminator-based system. Place a sample on one of four application-specific trays, then push the green Run button on the front of the imager to start the default protocol and image the sample using Image Lab.
Instrument Guide | 11
1 | Introduction
Four sample trays are available for the Gel Doc EZ imager. Each tray supports applications requiring a different set of stains and detection reagents, as shown in Table 3.
Table 3. Sample Tray Types and Detection Reagents
UV Tray Catalog #1708271
Ethidium bromide Fast Blast™ DNA
SYBR® Green Coomassie Blue
SYBR® Safe Copper stain SYBR® Gold
SYBR® Gold Zinc stain GelGreen
GelGreen Silver stain
GelRed
Flamingo™ fluorescent gel stain
Oriole™ fluorescent gel stain
Coomassie Fluor Orange
SYPRO Ruby
Krypton
White Light Tray Catalog
#1708272
stain
stain
Blue Tray Catalog
#1708273
SYBR® Green Stain-free gels
SYBR® Safe Stain-free blots
Stain-Free Tray Catalog
#1708274
12 | Gel Doc EZ Imaging System with Image Lab Software

Technical Specifications

Technical Specifications
Hardware Specifications
Image area 15 x 11.2 cm
Excitation source Trans UV B (302 nm peak)
Detector Charge-coupled device
Image pixel size 107.8 x 107.8 m (in microns)
Dynamic range 3.0 orders of magnitude
Pixel density 4,096 gray levels
Flat fielding Yes
Instrument size 43 x 28 x 38 cm
Instrument weight 7.3 kg
Connector descriptions USB-A connector is used for communication with a PC
USB-B connector is used for the instrument interface
Operating Ranges
Operating voltage 110/115/230/240 VAC Nominal
Operating temperature 10–28
Operating humidity < 70% noncondensing
Input voltage range 100–240 VAC
Input frequency range 50–60 Hz
Power 40 W
The Gel Doc EZ imager is for indoor laboratory use only.
Workflow automated execution
Workflow reproducibility 100% repeatability via recallable protocols, from gel activation
Autoexposure 2 user-defined modes (intense or faint bands)
C (21C recommended)
Equipment Ratings
Automation Capabilities
Controlled by a protocol via setup for gel activation, image capture, analysis, and reporting
and image capture to quantitative analysis and reports
Instrument Guide | 13
1 | Introduction

Image Lab Software Capabilities

Image Lab software runs customizable, automated protocols on the Gel Doc EZ system for routine gel and blot imaging and analysis.

Gel Documentation

Gel electrophoresis is a common way to separate, identify, and purify proteins or nucleic acids. The Gel Doc EZ system enables you to image and print gel and blot images for documentation in laboratory notebooks and to export images for publication or presentation.

Molecular Weight Assessment

Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and agarose gel electrophoresis are used to separate proteins or nucleic acids according to their size. Protein or nucleic acid molecular weight standards provide a reference for estimating the molecular weight of the proteins or nucleic acids being tested.

Quantitation

The components in a sample (bands) are quantitated to determine relative amounts of each component in a sample or to compare the amount of a sample component relative to a standard. Assessing the purity of a sample requires the quantitation of all components of a sample relative to each other. The results are expressed either as a percentage of all bands identified (band%) or as a percentage of all signals in the sample lane (lane%). Determining expression levels of a protein or nucleic acid requires quantitation among samples. Data can be reported as either relative values, if the quantity is unknown, or as an absolute value, if a standard of known quantity is present.

For More Information

For detailed information about Image Lab software, see the Image Lab Software User Guide.
14 | Gel Doc EZ Imaging System with Image Lab Software

2 Setting Up Gel Doc EZ

Configuring the Imager

Configure the Gel Doc EZ imager before using it with Image LabTM software. To do so, you must initialize the sample trays and correct the dark image.
Correcting the dark image reduces dark current noise generated from the charge-coupled device (CCD). Dark current noise is typical of all CCDs and is a result of the accumulation of charge in the absence of light.
To configure the imager
1. Verify that the Gel Doc EZ is connected to the computer running Image Lab software.
2. If a tray is in the imager, remove it and close the door.
3. Turn on the imager and start Image Lab software.
The setup wizard appears.
Instrument Guide | 15
2 | Setting Up Gel Doc EZ
4. Complete the steps in the wizard.
Note: You must complete all the steps in the setup wizard or the imager will not be usable. However, if you are not going to use a certain tray type, you can skip configuring that tray type and go to the next screen.
The first page of the setup wizard indicates that a new instrument has been found and displays the instrument serial number.
1 Click Next.
16 | Gel Doc EZ Imaging System with Image Lab Software
Loading...
+ 40 hidden pages