Real-Time PCR

CFX Duet Real-Time PCR System
Instrument Guide

Visit bio-rad.com/CFXDuet for more information.

CFX Duet Real-Time PCR Detection
SystemInstrument Guide
For Research Use Only

Bio-Rad Technical Support
The Bio-Rad Technical Support department in the U.S. is open Monday through Friday, 5:00 AM to
5:00 PM, Pacific Time.
Phone: 1-800-424-6723, option 2
Email: Support@bio-rad.com (U.S./Canada Only)
For technical assistance outside the U.S. and Canada, contact your local technical support office or click
the Contact us link at www.bio-rad.com.

NoticeNo part of this publication may be reproduced or transmitted in any form or by any means, electronic ormechanical, 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 guide is subject to change
without notice. Although prepared to ensure accuracy, Bio-Rad assumes no liability for errors oromissions, or for any damage resulting from the application or use of this information.
BIO-RAD, HARD-SHELL, and MICROSEAL are trademarks of Bio-Rad Laboratories, Inc. in certain
jurisdictions.
SYBR is a trademark of Thermo Fisher Scientific Inc. Bio-Rad Laboratories, Inc. is licensed to sell reagents
containing SYBR Green I for use in real-time PCR, for research purposes only.
All trademarks used herein are the property of their respective owner.
Copyright © 2021 by Bio-Rad Laboratories, Inc. All rights reserved.

Revision History

Revision History
Document
CFX Duet Real-Time PCR Detection

Date

Description of
ChangeDecember 2021

Initial Release

System Instrument Guide(Doc ID #10000148048)

Instrument Guide | iii

iv | CFX Duet Real-Time PCR System

Table of Contents
Revision History

Safety and Regulatory Complianceiii

9

Safety Warning Labels

9

Safety and Regulatory Compliance10

Safety Compliance

10

Electromagnetic Compatibility (EMC)11

EMC Warnings and Notes

11

Environment Requirements

13

Hazards

14

Biohazards

14

Chemical Hazards

15

Explosive or Flammability Hazards

15

Electrical Hazards

15

Transport

16

Battery

16

Warranty

16

Chapter 1 Introduction

17

Main Features

17

Performance Specifications

18

Sample Block Performance Specifications

18

Optical Detection Performance Specifications

19

Finding Out More

Chapter 3 Setting Up the CFX Duet Real-Time PCR Detection System
Site Requirements

19

21
21

Bench Space Requirements

21Power Requirements

22

System Overview

23

Front View

23

Instrument Guide | v

Table of Contents

Back View

25

Unpacking and Starting the Instrument

26

Packaged Instrument Specifications

26

Unpacking the Instrument

28

Connecting the Power Cables and Communication Cables

31

Starting the CFX Duet Real-Time PCR Detection System and Connecting to CFX Maestro

32

Removing the Shipping Screw

32

Loading the Sample Block

34

Shutting Down the CFX Duet Real-Time PCR Detection System

35

Chapter 4 Configuring the CFX Duet Real-Time PCR Detection System

37

Connecting to a Computer Running CFX Maestro Software

37

Renaming the CFX Duet Real-Time PCR Detection System

38

Viewing CFX Duet Real-Time PCR Detection System Total Run Hours and Total Cycles

39

Parameters and Ranges for Protocol Steps

40

Calibrating New Dyes

42

Appendix A Bio-Rad’s Real-Time PCR Detection Systems and CFX Maestro Software
Catalog Numbers
45
Appendix B Recommended Plastic Consumables

47

Plastic Consumables for the CFX Duet System

47

Plate Seals and Plate Sealer

47

Appendix C Maintenance and Troubleshooting

49

Cleaning and Maintaining the CFX Duet Real-Time PCR Detection System

49

Maintaining Sufficient Air Flow

55

Testing for Sufficient Air Flow

55

Fixing Insufficient Air Flow55

Replacing Fuses

56

Upgrading Software and Firmware on CFX Duet Real-Time PCR Systems

60

Shutting Down the CFX Duet Real-Time PCR Detection System

61

Returning the CFX Duet Real-Time PCR Detection System to Bio-Rad

62

Installing the Shipping Plate and Screw

Appendix D Bio-Rad Free and Open-Source Notices for PCR Products
Software Notices
M2Mqtt (paho.mqtt.m2mqtt)vi | CFX Duet Real-Time PCR System

62

65
66
66

Table of Contents

Standard Open License Text
EPL 1.0

Appendix E References

70
70

77

Instrument Guide | vii

Table of Contents

viii | CFX Duet Real-Time PCR SystemSafety and Regulatory Compliance
The CFX Duet Real-Time PCR Detection System (known in this guide as CFX Duet) heats and cools very
quickly during operation. Bio-Rad strongly recommends that you follow the safety specifications listed
throughout this guide.
Note: Use only Bio-Rad–supplied USB cables (catalog # 16007721) when using the CFX Duet
instrument.

Safety Warning Labels
Warning labels posted on the instrument and in this manual warn you about sources of injury or harm.
Table 1 defines each safety warning label.
Table 1. Meaning of safety warning labels

Icon

Meaning
Operating the CFX Duet instrument before reading this manual can constitute a personal
injury hazard. For safe use, do not operate this instrument in any manner unspecified in thismanual. Only qualified laboratory personnel trained in the safe use of electrical equipment
should operate this instrument. Always handle all components of the system with care andwith clean, dry hands.

Warning about handling biohazardous materials
When handling biohazardous samples, adhere to the recommended precautions andguidelines and comply with any local guidelines specific to your laboratory and location.

Warning about risk of burning
A thermal cycler generates enough heat to cause serious burns. Wear safety goggles orother eye protection at all times during operation. Always allow the sample block to return toidle temperature before opening the lid and removing samples. Always allow maximum
clearance to avoid accidental skin burns.

Warning about risk of explosion
The sample blocks can become hot enough during the course of normal operation to cause
liquids to boil and explode.

Instrument Guide | 9

Safety and Regulatory Compliance

Safety and Regulatory Compliance
Safety Compliance
The CFX Duet instrument has been tested and found to be in compliance with all applicablerequirements of the following safety and electromagnetic standards:n

IEC 61010-1:2010 Safety requirements for electrical equipment for measurement, control, andlaboratory use, Part 1: General requirementsn

IEC 61010-2-010:2019 Safety requirements for electrical equipment for measurement, control
and laboratory use — Part 2-010: Particular requirements for laboratory equipment for the heatingof materialsn

IEC 61010-2-081:2019 Safety requirements for electrical equipment for measurement, control
and laboratory use — Part 2-081: Particular requirements for automatic and semi-automatic
laboratory equipment for analysis and other purposesn

CAN/CSA-C22.2 NO. 61010-1-12:2018 Safety requirements for electrical equipment formeasurement, control, and laboratory use, Part 1: General Requirementsn

CAN/CSA-C22.2 NO. 61010-2-010:19 Safety requirements for electrical equipment formeasurement, control, and laboratory use, Part 2-010: Particular requirements for laboratory
equipment for the heating of materialsn

CAN/CSA-C22.2 NO. 61010-2-081:19 Safety requirements for electrical equipment formeasurement, control, and laboratory use, Part 2-081: Particular requirements for automatic andsemi-automatic laboratory equipment for analysis and other purposesn

EN 61010-1:2010 Safety requirements for electrical equipment for measurement, control, andlaboratory use, Part 1: General requirementsn

EN 61010-2-010:2014 Safety requirements for electrical equipment for measurement, control andlaboratory use — Part 2-010: Particular requirements for laboratory equipment for the heating ofmaterialsn

EN 61010-2-081:2015 Safety requirements for electrical equipment for measurement, control andlaboratory use — Part 2-081: Particular requirements for automatic and semi-automatic laboratory
equipment for analysis and other purposes

10 | CFX Duet Real-Time PCR SystemSafety and Regulatory Compliancen

UL 61010-1:2012 Safety requirements for electrical equipment for measurement, control andlaboratory use — Part 1: General Requirementsn

UL 61010-2-010:2019 Safety requirements for electrical equipment for measurement, control andlaboratory use — Part 2-010: Particular requirements for laboratory equipment for the heating ofmaterialsn

UL 61010-2-081:2019 Safety requirements for electrical equipment for measurement, control andlaboratory use — Part 2-081: Particular requirements for automatic and semi-automatic laboratory
equipment for analysis and other purposesElectromagnetic Compatibility (EMC)
The CFX Duet instrument has been tested and found to be in compliance with all applicablerequirements of the following electromagnetic compatibility standards:n

IEC 61326-1:2012 Electrical equipment for measurement, control and laboratory use — EMC
requirements — Part 1: General requirements. Tested as a Class A devicen

EN 61326-1:2013 Electrical equipment for measurement, control and laboratory use — EMC
requirements — Part 1: General requirements. Tested as a Class A devicen

FCC Part 15, Subpart B, Sections 15.107 and 15.109. Tested as a Class A digital devicen

CAN ICES-003v6:2019 Interference-causing equipment standard, information technology
equipment (including digital apparatus) — Limits and methods of measurement. Tested to Class
A limits

EMC Warnings and NotesnWarning: Changes or modifications to this unit, not expressly approved by Bio-Rad, could void
the user’s authority to operate the equipment.

nNote: This equipment has been tested and found to comply with the limits for a Class A digital
device, pursuant to part 15 of the FCC Rules. These limits are designed to provide reasonable
protection against harmful interference when the equipment is operated in a commercial
environment. This equipment generates, uses, and can radiate radio frequency energy and, if not
installed and used in accordance with the instruction manual, may cause harmful interference toradio communications. Operation of this equipment in a residential area is likely to cause harmfulinterference, in which case the user will be required to correct the interference at their own
expense.

nNote regarding FCC compliance: Although this instrument has been tested and found to comply
with Part 15, Subpart B of the FCC Rules for a Class A digital device, please note that thisInstrument Guide | 11Safety and Regulatory Compliancecompliance is voluntary, for the instrument qualifies as an “exempted device” under 47 CFR
15.103(c), in regard to the cited FCC regulations in effect at the time of manufacture.
nNote regarding cables: This instrument was tested for EMC compliance using specially designed
USB cables, which are supplied with the instrument. These cables, or Bio-Rad authorized
replacements, must be used with this instrument to ensure continued compliance with the EMC
emissions limits.

12 | CFX Duet Real-Time PCR SystemSafety and Regulatory Compliance

Environment Requirements
The CFX Duet has been designed to be safely operated under the environmental conditions listed in
the following table.
Table 2. environment requirements

Parameter

Specification

Environment

Indoor use only

Operating altitude

Up to 2,000 meters above sea level

Ambient room temperature

15–31°C*

Transport and storage temperature–20° to 60°C**–4 to 140°F

Relative humidity

20% to 80% (noncondensing)***Operating power

100 to 240 VAC ±10%, 50/60 Hz, 850 W Max

Mains supply voltage fluctuation±10%

Maximum power usage<850 watts

Fuses

10 A, 250 V, 5 x 20 mm, fast blow (qty. 2)

Overvoltage category

II

Pollution degree

2*Operating the instrument outside of this temperature range may not meet performance
specifications. A room temperature between 5–40°C is considered safe.
**Store and transport the instrument in its shipping container to meet these temperature conditions.
***Operating the instrument at 4⁰C should be limited to 18 hours at these conditions. Holds at 4⁰C canbe performed for up to 72 hours if humidity is less than 60% (noncondensing).

Instrument Guide | 13

Safety and Regulatory ComplianceHazards
The CFX Duet instrument is designed to operate safely when used in the manner prescribed by the
manufacturer. If the system or any of its associated components is used in a manner not specified by
the manufacturer, the inherent protection provided by the instrument may be impaired. Bio-Rad is notliable for any injury or damage caused by the use of this equipment in any unspecified manner, or bymodifications to the instrument not performed by Bio-Rad or an authorized agent. Service of the CFX
Duet instrument should be performed only by trained Bio-Rad personnel.

Biohazards
The CFX Duet instrument is a laboratory product. However, if biohazardous samples are present,
adhere to the following guidelines and comply with any local guidelines specific to your laboratory
and location.
Note: No biohazardous substances are exhausted during normal operations of this instrument.

General Precautionsn

Always wear laboratory coat, laboratory gloves, and safety glasses with side shields or goggles.

n

Keep your hands away from your mouth, nose, and eyes.

nCompletely protect any cut or abrasion before working with potentially infectious materials.

nWash your hands thoroughly with soap and water after working with any potentially infectious
material before leaving the laboratory.

n

Remove wristwatches and jewelry before working at the bench.

nStore all infectious or potentially infectious material in unbreakable leak-proof containers.

nBefore leaving the laboratory, remove protective clothing.

n

Do not use a gloved hand to write, answer the telephone, turn on a light switch, or touch anythingthat other people may touch without gloves.

n

Change gloves frequently. Remove gloves immediately when they are visibly contaminated.

n

Do not expose materials that cannot be properly decontaminated to potentially infectious material.

nUpon completion of an operation involving biohazardous material, decontaminate the work areawith an appropriate disinfectant (for example, a 1:10 dilution of household bleach).

14 | CFX Duet Real-Time PCR System

Hazards

Surface Decontamination
WARNING! To prevent electrical shock, always turn off and unplug the instrument
prior to performing decontamination procedures.

The following areas can be cleaned with any hospital-grade bactericide, virucide, or fungicide
disinfectant:nOuter lid and chassisn

Inner sample block surface and sample block wells

To prepare and apply the disinfectant, refer to the instructions provided by the product manufacturer.
Always rinse the sample block and sample block wells several times with water after applying adisinfectant. Thoroughly dry the sample block and sample block wells after rinsing with water.
Important: Do not use abrasive or corrosive detergents or strong alkaline solutions. These agentscan scratch surfaces and damage the sample block, resulting in loss of precise thermal control.

Disposal of Biohazardous Material
Dispose of the following potentially contaminated materials in accordance with laboratory local,
regional, and national regulations:nClinical samples

nReagents

n

Used reaction vessels or other consumables that may be contaminatedChemical Hazards
The CFX Duet instrument contains no potentially hazardous chemical materials.

Explosive or Flammability Hazards
The CFX Duet instrument poses no uncommon hazard related to flammability or explosion when used
in a proper manner as specified by Bio-Rad Laboratories.

Electrical Hazards
The CFX Duet instrument poses no uncommon electrical hazard to operators if installed and operated
properly without physical modification and connected to a power source of proper specification.

Instrument Guide | 15

Safety and Regulatory ComplianceTransport
Before moving or shipping the CFX Duet instrument, decontamination procedures must be performed.
Always move or ship the system in a separate container in the Bio-Rad-supplied packaging material,
which protects the system from damage.
For information about transporting the system and to request the appropriate packaging material,
contact your local Bio-Rad office.

Battery
The CFX Duet instrument uses one 3 V lithium-metal coin cell battery to maintain time settings in theevent of AC power loss. If the time does not remain set after the unit is turned off, it may be anindication that the batteries are getting weak.
WARNING! Do not attempt to change the batteries. They are not user serviceable.
Instead, contact Bio-Rad Technical Support for assistance.

For the State of California, USA onlynPerchlorate material — Lithium batteries contain perchlorate material; special handling may
apply. See www.dtsc.ca.gov/hazardouswaste/perchlorate.

Warranty
The CFX Duet instrument and its associated accessories are covered by a standard Bio-Rad warranty.
Contact your local Bio-Rad office for the details of the warranty.

16 | CFX Duet Real-Time PCR System

Chapter 1 Introduction
The CFX Duet Real-Time PCR Detection System (known in this guide as CFX Duet) is a real-time
PCR system with an integrated 96-well sample block.
When used with the CFX Maestro Software (known in this guide as CFX Maestro), CFX Duet becomesa powerful real-time PCR detection system for researchers who run polymerase chain reaction (PCR)
for sequencing, cloning, gene expression studies, mutagenesis, and many other applications.
This guide explains how to set up CFX Duet to perform sensitive quantitative PCR.
Important: The CFX Duet instrument requires the latest version of CFX Maestro to perform PCR
runs. You will not be able to perform PCR runs on the CFX Duet instrument without CFX Maestro.
For instructions on how to use CFX Duet to perform PCR experiments and analyze PCR data, refer tothe CFX Maestro User Guide.

Main Features
The main features of CFX Duet include:n

USB ports enable direct connectivity to a computer running CFX MaestronOptical shuttle technology that does not require ROX or another normalizernProgrammable temperature gradient to quickly and easily identify optimal annealing
temperaturesn

Uniform thermal performance across the reaction blockn

Two channel optics for simultaneous dual target detectionn

FRET mode for protein characterization workflows.

Instrument Guide | 17

Chapter 1 Introduction

Performance Specifications
CFX Duet components provide sensitive detection for precise quantification and target discrimination.
The tables in this section list the sample block and optical detection performance specifications for the
CFX Duet instrument.

Sample Block Performance Specifications
Table 3. CFX Duet Real-Time PCR Detection System sample block specifications

Item

Specification

Volume1–50 µl (10–50 µl Recommended)

Maximum ramp rate

5°C/sec

Average ramp rate

3.3°C/sec

Lid temperature

30–110°C

Heating and cooling method

PeltierTemperature range

4–100°C

Increment-10°C to 10°C/cycle

Gradient
Operational range

30–100°C

Programmable span

1–24°C

Temperature accuracy±0.2°C of programmed target at 90°C

Temperature uniformity±0.3°C well-to-well within 10 sec of arrival at 90°C

18 | CFX Duet Real-Time PCR System

Finding Out More

Optical Detection Performance Specifications
Table 4. CFX Duet Real-Time PCR Detection System optical detection specifications

Item

Specification

Excitation

3 filtered LEDs

Detection

3 filtered photodiodes

Range of excitation/emission wavelengths
Multiplex analysis

450–580 nm
2 targets per well

Scan time
All channels

12 secSingle-channel fast scan

3 sec

FRET

Yes

Sensitivity

Detects 1 copy of target sequence inhuman genomic DNA

Dynamic range

10 orders of magnitude

Finding Out More
You can access the CFX Maestro User Guide, which describes the software’s full features, in the Help
Menu of the CFX Maestro Home Window. You can also access the Maestro User Guide by logging
onto bio-rad.com and searching for catalog #10000126764.
Note: Click the Bio-Rad logo in the upper right corner of any CFX Maestro window to launch
Bio-Rad’s website. This site includes links to technical notes, manuals, product information, andtechnical support. This site also provides many technical resources on a wide variety of methods
and applications related to PCR, real-time PCR, and gene expression.
The CFX Duet Real-Time PCR Detection System web site (bio-rad.com/CFXDuet) provides access totechnical notes, manuals, product information, and technical support. This site also provides manytechnical resources on a wide variety of methods and applications related to real-time PCR.

Instrument Guide | 19

20 | CFX Duet Real-Time PCR System

Chapter 3 Setting Up the CFX Duet Real-TimePCR Detection System
This chapter explains how to set up the CFX Duet instrument at your site.
Tip: Before setting up the CFX Duet instrument, familiarize yourself with the system, its ports, andaccessories.

Site Requirements
The tables in this section list the bench space and power requirements necessary to successfully
install and use the CFX Duet instrument. For weight and dimensions of the packaged instrument, see
Packaged Instrument Specifications on page 26.
Note: Install your CFX Duet instrument on a flat, dry surface with sufficient cool airflow for it to run
properly.

Bench Space Requirements
Caution: The CFX Duet instrument requires at least 10 cm behind and at
least 5 cm along one side or the top to allow exhaust air to flow properly.

Additionally, ensure you provide sufficient space around the system to disconnect the power cordor to access the power switch in case of an emergency.
Note: While setting up the CFX Duet instrument, ensure that you have sufficient space near thesystem for the computer running CFX Maestro.

Instrument Guide | 21

Chapter 3 Setting Up the CFX Duet Real-Time PCR Detection System

Table 5. CFX Duet Real-Time PCR Detection System bench space requirements

Item

Specification

Dimensions

W: 13 in; 33 cm
D: 22 in; 56 cm
H: 14 in; 36 cm

Weight48 lb; 22 kg

Power Requirements
Power to the CFX Duet instrument must be stable and within specifications to ensure proper
operation. The power cord connected to the power inlet port must be rated for 10 amps or more.
Table 6. CFX Duet Real-Time PCR Detection System power requirements

Item

Specification

Fuses

10 A, 250 V, 5 x 20 mm, fast blow (qty. 2)

Number of power sockets

n One socket for the CFX Duet instrumentn One socket for the computer running CFX Maestro

22 | CFX Duet Real-Time PCR System

System Overview

System Overview
The illustrations in this section display the main components of the CFX Duet instrument.

Front View

LEGEND
1. Sample block

2. Retractable lid (partially retracted)

3. Air intake vents

4. LED system status light

Detailsn

Sample block — holds the 96-well plate.

nRetractable lid — provides access to the sample block.
Note: The CFX Duet instrument does not have a manual open/close button on the lid. To
open or close the lid, use the lid control in CFX Maestro.

n

LED system status light — indicates system status:oGreen (constant) — run is in progress.

oGreen (flashing) — run is paused.

oBlue (flashing) — run has completed (flashes until lid has opened).

oBlue (flashes for ten seconds) — system is in use (indicates that the instrument is selected in
CFX Maestro).

Instrument Guide | 23

Chapter 3 Setting Up the CFX Duet Real-Time PCR Detection Systemo

Red (flashing) — critical system error.

oWhite — system is idle (not executing a run).
Important: The system will become idle (indicated by a white LED) if the CFX Duet
instrument disconnects from CFX Maestro.

on

OFF — system is shut down.

Air intake vents — allow the system to heat and cool quickly.
Important: Keep all air vents clean and clear of obstruction. Do not insert
any object in the air vents at any time. Fans or other internal moving parts
may collide with the object and cause personal injury or damage theinstrument. Conductive objects could contact internal circuitry and cause
damage to the instrument.

24 | CFX Duet Real-Time PCR System

System Overview

Back View

LEGEND
1. Power input

2. Power switch

3. Fuses

4. Cooling vents

5. USB Type B port

Detailsn

Power input — connects to AC power.

nPower switch — turns the CFX Duet instrument power on or off.

nFuses — provides access to the fuses.

n

Cooling vents — cool the instrument.
Important: Do not block the cooling vents. For optimal operation, ensure that air can circulate
behind the instrument.

n

USB Type B port — connects the CFX Duet instrument to a computer running CFX Maestro.

Instrument Guide | 25

Chapter 3 Setting Up the CFX Duet Real-Time PCR Detection System

Unpacking and Starting the Instrument
This section explains how to properly unpack and start the CFX Duet instrument. Read this section
before you begin.
Caution: The packaged CFX Duet instrument weighs approximately 57 lb (26 kg). Bio-Rad
strongly recommends that you use a pallet jack to move the instrument from the shipping dock to
the laboratory.
Important: Use proper lifting techniques when moving and lifting the instrument
to prevent damage to the instrument and personal injury. Bio-Rad recommends
that two or more people lift the instrument.

Packaged Instrument Specifications
Table 7 lists the specifications of the packaged CFX Duet instrument. For information about bench
space specifications, see Site Requirements on page 21.
Table 7. Specifications for the packaged CFX Duet Real-Time PCR Detection System

Parameter

Specification

Weight57 lb; 26 kg

Dimensions

Depth: 28 in; 71 cm
Width: 19 in; 48 cm
Height: 19 in; 48 cm

26 | CFX Duet Real-Time PCR System

Unpacking and Starting the Instrument

The image below depicts the CFX Duet instrument inside its packaging.

Instrument Guide | 27

Chapter 3 Setting Up the CFX Duet Real-Time PCR Detection System

Unpacking the Instrument
Install the CFX Duet instrument on a flat, dry surface with sufficient cool airflow to run properly. The
AC power cord is approximately 5 ft. in length. Ensure you have sufficient access to the power outlet
before you begin.
Note: Ensure that there is enough clear space on the lab bench for your instrument plus
additional space to maneuver it while installing cables and accessories. Additionally, ensure thatyou have adequate space on the lab bench for the computer running CFX Maestro.
Caution: When handling the instrument, do not grip the retractable lid. To lift the instrument,
position your hands underneath the right and left sides of the instrument.

To unpack the system
1.

Ensure that the path from the shipping dock to the laboratory is clear of obstacles and canaccommodate the packaged CFX Duet instrument and a pallet jack.

2.

Using a pallet jack, move the instrument container from the shipping dock to the laboratory.
Important: Bio-Rad strongly recommends that you do NOT use a hand truck to move thepackaged instrument.

3.

Release the instrument container next to the lab bench.

4.

Using two people, lift and place container onto the lab bench.

5.

With scissors or a box cutter, cut and remove the exterior straps that secure the container top to
the base.
Tip: The exterior straps are very tight. Hold the straps securely while cutting to prevent injury.

6.

Remove the plastic clips on the bottom corners by firmly pinching the tabs together and pullingthe clips straight out.

28 | CFX Duet Real-Time PCR System

Unpacking and Starting the Instrument

7.

Lift the large container top up and out of the base and place it to the side.

8.

Remove the top foam insert and place it to the side.

Instrument Guide | 29

Chapter 3 Setting Up the CFX Duet Real-Time PCR Detection System

9.

Standing in front of the instrument, fold down the front of the cardboard base.

10. While another person holds the base, grasp the cardboard tray and slide the instrumentcompletely out of the base.

11. Using the lift point on one side of the instrument, gently lift the instrument, remove the foam
inserts, and place them to the side.
12. Using two people, carefully lift the instrument and remove the cardboard tray.
13. Slide the plastic cover toward the back of the instrument to remove.

30 | CFX Duet Real-Time PCR System

Unpacking and Starting the Instrument

14. Inspect the instrument for any damage.
Important: If you observe any damage to the system, do not continue.
Instead, contact Bio-Rad Customer Support.

Connecting the Power Cables and Communication Cables
After you unpack the CFX Duet instrument and install it on the lab bench, you will need to connect thepower cord to a power source and a USB Type B cable to a computer running CFX Maestro. Thissection explains how to connect the power cord and connect to CFX Maestro.
Tip: Before connecting the cables, familiarize yourself with the CFX Duet instrument, its accessory
kit, and power switches.
Important: Ensure you have enough room on the lab bench to reach the power switch on theback of the system after connecting the power cord and USB Type B cable. Use only the Bio-Radsupplied power cord and USB Type B cable.

To connect the CFX Duet instrument power and communication cables
1.

Locate the accessory kit that shipped with the CFX Duet instrument.

2.

Remove the AC power cord and USB cable from the accessory kit.
Tip: Store the packing material for future use. If any item is missing or damaged, contact yourlocal Bio-Rad office.

3.

Insert one end of the AC power cord into the power inlet port on back of the system.

4.

Insert the other end of the AC power cord into an available grounded, surge-protected AC outlet.

5.

To connect the CFX Duet instrument to a computer running CFX Maestro, insert the male end of
the USB Type B cable into the USB Type B port located on the back of the system. Connect theother end of the USB Type B cable into a USB port on the computer running CFX Maestro.

Important: You must install CFX Maestro before you connect the computer running CFX Maestro
to the CFX Duet instrument.

Instrument Guide | 31

Chapter 3 Setting Up the CFX Duet Real-Time PCR Detection System

Starting the CFX Duet Real-Time PCR Detection System andConnecting to CFX Maestro
Important: CFX Maestro must be installed on a computer connected to the CFX Duet instrument
in order for the software to recognize the instrument. For instructions on how to install and log into
CFX Maestro, refer to the CFX Maestro user guide.

To start the CFX Duet Real-Time PCR Detection System
1.

Turn on the computer running CFX Maestro.

2.

Press the power switch on the back of the instrument to start the CFX Duet instrument.

3.

The system performs a series of initialization tests. The CFX Duet instrument then appears in the
CFX Maestro Detected Instruments pane.
Note: If the CFX Duet instrument does not appear in the Detected Instrument pane of CFX
Maestro, verify that the USB cable is properly installed. To reinstall drivers, on the Home
window in CFX Maestro select Tools > Reinstall Instrument Drivers.

Removing the Shipping Screw
The CFX Duet instrument ships with a red shipping screw inserted in the inner lid to stabilize theoptical shuttle during shipping. You must use CFX Maestro to prepare the instrument for screw
removal before unscrewing the shipping screw from the CFX Duet instrument.

To remove the shipping screw
1.

Locate the beige shipping screw plug that ships with the CFX Duet instrument.
Tip: This might be located in a plastic sheath taped to the front or side of the instrument.

2.

Ensure the power cord is properly inserted into the power input on the back of the instrument.

3.

If you have not yet done so, insert the other end of the power cord into an available grounded,
surge-protected AC outlet.

4.

Press the power switch on the back of the instrument to start the CFX Duet instrument.

5.

In CFX Maestro, confirm that the CFX Duet instrument icon appears in the Detected Instruments
pane.
Note: If the CFX Duet does not appear in the Detected Instruments pane of CFX Maestro, seeConnecting the Power Cables and Communication Cables on page 31.

6.

In the Detected Instruments pane of CFX Maestro, right-click the CFX Duet and select Properties.

7.

Select the Shipping Screw tab and click Remove Shipping Screw.

32 | CFX Duet Real-Time PCR System

Unpacking and Starting the Instrument

8.

Remove the shipping screw from the side of the CFX Duet instrument lid by unscrewing it counterclockwise.
Important: You must reinsert the shipping screw should you need to return the instrument for
any reason. Save the screw in a safe and accessible place.
Caution: Do not insert the shipping screw or any other object in the shipping
screw hole while instrument is operating. Internal moving parts might collidewith the object, which can cause personal injury or damage the instrument.

9.

In CFX Maestro, click Screw Removed to confirm that the shipping screw is removed.

10. Click Open Lid.
11. Remove the shipping plate from the CFX Duet instrument that protects the sample block.
12. Click Close Lid.
13. Close the Properties dialog box.

Instrument Guide | 33

Chapter 3 Setting Up the CFX Duet Real-Time PCR Detection System

Loading the Sample Block
Bio-Rad strongly recommends that you use only low-profile plates and flat-cap tubes with the CFX
Duet instrument. Using high-profile plates can crush tubes. Using domed-cap tubes can negatively
affect plate reads.
For a list of plates and tubes compatible with the CFX Duet, visit us at www.bio-rad.com/cfxopus orcontact your local Bio-Rad Sales representative.
To ensure uniform heating and cooling of samples, reaction vessels must be in complete contact withthe sample block. To ensure adequate contact, do the following:nConfirm that the sample block is clean before loading samples.

nFirmly press the individual tubes, tube strips, or microplates into the block wells.

nWhen using one or a few tubes, use the tube frame or load at least one empty tube in each cornerof the block to ensure the lid exerts even pressure on individual tubes.

Loading Plates, Tubes and Tube Strips into the Sample Block
Caution: Never run a sample with a lid or seal that is open, loose, punctured, orotherwise damaged. Doing so will increase the likelihood of a rupture, which could
cause injury or contaminate the instrument.
Important: When running the CFX Duet instrument, always balance the tube strips or add capped
tubes to the corner wells to ensure the heated lid applies even pressure across the block.

To load plates into the sample block
1.

2.

To open the motorized lid, do one of the following in CFX Maestro:n

In the Detected Instruments pane, click Open Lid.

n

In the Detected Instruments pane, right-click on the instrument and click Open Lid.

n

On the Start Run tab of the Run Setup pane, click Open Lid.

Place the microplate, individual tubes, or tube strips with sealed lids in the block.
Important: Ensure that the tubes are completely sealed to prevent leakage.
Tip: For optimal results, load sample volumes of 10–50 µl.

3.

For accurate data analysis, verify that the orientation of samples in the block is exactly the sameas the orientation of the well contents in CFX Maestro.

4.

To close the motorized lid, do one of the following:n

In the Detected Instruments pane of CFX Maestro, click Close Lid.

34 | CFX Duet Real-Time PCR System

Shutting Down the CFX Duet Real-Time PCR Detection Systemn

On the Start Run tab of the Run Setup pane of CFX Maestro, click Close Lid.

n

In the Detected Instruments pane of CFX Maestro, right click the instrument and click Close
Lid.

Important: Ensure that nothing blocks the lid when it closes. Although there is a safetymechanism to prevent the lid from closing if it senses an obstruction, do not place anything in
the way of the lid before closing.

Shutting Down the CFX Duet Real-Time PCR Detection
SystemFollow these instructions to safely and completely shut down the CFX Duet instrument.

To shut down the CFX Duet instrument
1.

Ensure that no protocol is running and the instrument is no longer in use.

2.

In the CFX Maestro Detected Instruments pane, select the CFX Duet instrument.

3.

Click Open Lid.

4.

Remove the sample plate located on the block of the CFX Duet instrument.

5.

Click Close Lid.

6.

Press the power switch on the back panel of the instrument to power down the system.

Instrument Guide | 35

Chapter 3 Setting Up the CFX Duet Real-Time PCR Detection System

36 | CFX Duet Real-Time PCR System

Chapter 4 Configuring the CFX Duet Real-TimePCR Detection System
After successfully setting up the CFX Duet instrument and installing CFX Maestro, you can configure
CFX Maestro (based on the your site's requirements) to perform the following:nConnect the CFX Duet instrument to a computer running CFX Maestron

Set the CFX Duet instrument’s time zone and local timenRename the CFX Duet instrumentnView the CFX Duet instrument's total number of PCR hours and cycles since it was last connected
to CFX Maestron

Set protocol parameters (such as ramp rate, gradient, and temperature)nCalibrate dyes

For more detailed information on how to perform these tasks, refer to the CFX Maestro User Guide.

Connecting to a Computer Running CFX Maestro Software
During installation, the CFX Maestro software installer automatically installs the instrument drivers
onto the computer. When you start the software, CFX Maestro detects the connected instruments.
Important: You must disconnect the CFX Duet instrument from the CFX Maestro computer before
you install the software. You do not need to turn off the instrument during the software installation.

To detect connected instruments
1.

If you have not yet done so, insert the square (male) end of the supplied USB Type B cable into
the USB Type B port located on the back of the CFX Duet instrument.

2.

Insert the other (port) end into a USB port on the CFX Maestro computer.

3.

If the instrument is not already running, press the power switch on the back of the instrument toturn it on.

4.

Start CFX Maestro.

Instrument Guide | 37

Chapter 4 Configuring the CFX Duet Real-Time PCR Detection System

The software automatically detects the connected instrument and displays its name in the Detected
Instruments pane in the Home window.

Note: If the CFX Duet instrument does not appear in the Detected Instruments pane, verify that theUSB cable is properly installed. To reinstall drivers, select Tools > Reinstall Instrument Drivers in
the CFX Maestro software Home window.

Renaming the CFX Duet Real-Time PCR Detection System
You can update the name of the CFX Duet instrument in CFX Maestro so that the instrument is moreeasily identifiable when performing PCR runs, analyses, and in run reports.

To rename the CFX Duet instrument:
1.

Start CFX Maestro.

2.

In the Detected Instruments pane, right-click the target instrument and select Properties on themenu that appears.

3.

The CFX Duet Instrument Properties dialog box appears.

4.

In the Properties tab, type a name in the Rename box at the top of the Properties tab and click
Rename.

5.

The new name appears in the Nickname row in the Properties tab as well as in the Instrument
Properties header bar and the Detected Instruments pane.

38 | CFX Duet Real-Time PCR System

Viewing CFX Duet Real-Time PCR Detection System Total Run Hours and Total Cycles

Viewing CFX Duet Real-Time PCR Detection System Total Run
Hours and Total Cycles
CFX Maestro allows for you to view the Total Run Hours and Total Cycles that the CFX Duet
instrument has performed since it was last connected to CFX Maestro.

To view the Total Run Hours and Total Cyclesu

In CFX Maestro, right-click on the CFX Duet instrument in the Detected Instruments pane, andselect Properties to display the Properties dialog. The Total Run Hours and Total Cycles aredefined as follows:n

Total Run Hours at Last Connection – the number of hours that the CFX Duet instrument
performed PCR cycles.

nTotal Cycles at Last Connection – the total number of cycles the CFX Duet instrument has
accumulated.

Total Run Hours and Total Cycles are updated each time the CFX Duet instrument connects to CFX
Maestro.

Instrument Guide | 39

Chapter 4 Configuring the CFX Duet Real-Time PCR Detection System

Total Run Hours and Total Cycles in the CFX Duet Instrument Properties tab:Parameters and Ranges for Protocol Steps
Create, edit, and run CFX Duet protocols in CFX Maestro. To create, edit, or run a protocol, refer to the
CFX Maestro User Guide.
Use the information in Table 8 on page 41 to modify the default settings for the steps in your protocol.

Temperature Steps
The target temperature is a value between 4.0 and 100.0°C, set in tenths of a degree. The system
ramps up to this temperature and holds that value for a specified amount of time (the hold time).

Gradient Steps
The gradient range is the difference between the lower and upper temperatures in a gradient step.
The maximum allowed range is 24°C. The lower temperature is a value between 30.0 and 99.0°C, set
in tenths of a degree. The maximum upper temperature is 100°C. The thermal cycler ramps up to thetarget temperature gradient across the block and holds that temperature for a specified hold time.
Important: CFX Maestro calculates the gradient value. When you enter a value in the gradientcalculator’s top and bottom fields in CFX Maestro, the software automatically calculates andassigns the temperatures for the remaining fields. When you enter a temperature in any field
between the top and bottom field, the software automatically calculates the remaining fields. You
cannot manually enter a temperature value in every field.

40 | CFX Duet Real-Time PCR System

Parameters and Ranges for Protocol Steps

For more information on how to enter gradient steps in CFX Maestro, refer to the CFX Maestro User
Guide.
Table 8. Parameters and ranges for protocol steps

Parameter

Range

Description

Ramp rate

0.1–5°C per sec

Instructs the thermal cycler toramp to the target temperature at
the specified rate in that step.
Available only to temperature
steps.

Increment

A number from –10.0 to 10.0°C per cycle in tenths

Instructs the thermal cycler toof a degreechange the target temperature of a
step with each cycle, where apositive number increases thetemperature and a negative
number decreases thetemperature.
Available only to temperature
steps that are in a GOTO loop.

ExtendA time from –60 to 60 sec per cycle

Instructs the thermal cycler toextend the hold time with eachcycle. A positive number
increases the hold time and anegative number decreases thehold time.
Available to both temperature andgradient steps that are in a GOTO
loop.

Beep

(No parameters)

Instructs the thermal cycler tobeep to signal that the thermal
cycler has reached the target
temperature for that step.
Available only to temperature
steps.

Plate read(No parameters)

Instructs the thermal cycler to add
a plate read to the selected step.
Available to both temperature andgradient steps.

Instrument Guide | 41

Chapter 4 Configuring the CFX Duet Real-Time PCR Detection System

Calibrating New Dyes
The CFX Duet instrument is factory calibrated for commonly used fluorophores in white-well and clearwell plates. Table 9 lists the fluorophores and channel for which each instrument is calibrated.
Note: The CFX Duet instrument also includes a channel dedicated to FRET chemistry. This
channel does not require calibration for specific dyes.
Important: If you conduct a user-defined calibration of a dye that was factory calibrated, theinstrument uses the user-defined calibration instead of the factory calibration. You must use CFX
Maestro to conduct a user-defined calibration.
Table 9. Factory calibrated fluorophores and channels

ChannelExcitation,nm

Detection,
nm

FAM, SYBR® Green I1

450–490515–530

VIC, HEX, CAL Fluor Gold 540, Cal Fluor Orange 560

2515–535

560–580

450–490560-580

Fluorophores

FRET Chemistry (Not Factory-Calibrated)
Non-factory calibrated color

42 | CFX Duet Real-Time PCR System

FRETCalibrating New Dyes

To calibrate new dyes for the CFX Duet instrument
1.

Start CFX Maestro.

2.

In the Home window of CFX Maestro, select the CFX Duet instrument in the Detected Instruments
pane.

3.

Select Tools > Dye Calibration Wizard to open the Dye Calibration wizard.

Fluorophores already calibrated for the target instrument appear in the Calibrated Fluorophores
table.
4.

In the Calibrate New or Existing Fluorophores section, select the fluorophore to calibrate from thedropdown list.
If the fluorophore name is not included in the list, type its name in the text box to add it to the list.
Important: Be careful when naming custom calibrated fluorophores. If you create a custom
dye calibration for a fluorophore with the same name as a factory calibrated fluorophore, thecustom fluorophore (not the factory calibrated fluorophore) will be the one used by the
instrument during the runs.

5.

Select the plate type for the fluorophore.
If the plate type is not included in the list, type the name in the text box to add it to the list.

6.

Select a channel for the fluorophore.

Instrument Guide | 43

Chapter 4 Configuring the CFX Duet Real-Time PCR Detection System

7.

Select a plate column for the fluorophore.

8.

(Optional) Type a color to associate with the fluorophore.

9.

Click Add to Plate to add the fluorophore.

10. (Optional) Repeat steps to add each fluorophore you plan to calibrate for the plate.
11. When you finish adding fluorophores, click View Plate to open the Pure Dye Plate Display
window.
Use this window as a guide for loading dyes into the plate.
12. Prepare a CFX Duet well plate for dye calibration:a.

Pipette dye solution into each well, following the pattern shown in the Pure Dye Plate
Display.

b.

For each fluorophore, fill four wells with 50 µl of 300 nM dye solution. Notice that at least half
the plate contains blank wells.

c.

Seal the plate using the sealing method you will use in your experiment.

13. Place the calibration plate in the block and close the lid.
14. In the Dye Calibration wizard, click Calibrate and then OK to confirm that the plate is in the block.
15. When CFX Maestro completes the calibration run, a dialog box appears. Click Yes to finish
calibration and open the Dye Calibration Viewer.
16. Click OK to close the window.

To view Calibrated Dyes:
1.

In the Detected Instruments Pane, right-click the CFX Duet instrument and select Properties.

2.

In the Instrument Properties Window, click the Calibrated Dyes tab.

To see detailed information about a calibration, click its Info button in the Detail column.

44 | CFX Duet Real-Time PCR System

Appendix A Bio-Rad’s Real-Time PCR Detection
Systems and CFX Maestro Software Catalog
Numbers
This appendix lists the catalog numbers for Bio-Rad’s CFX Duet Real-Time PCR Detection System,
CFX Maestro Software, and accessories.
Table 10. Catalog numbers for Bio-Rad’s CFX Duet, accessories, and CFX Maestro Software

Catalog Number

Description

Instruments
12016265

CFX Duet Real-Time PCR System

CFX Maestro Software and accessories
12013758

CFX Maestro Software 2.3

12012832

CFX Maestro Software SE

12012833

CFX Maestro Software, Russian Edition

12012834

CFX Maestro Software, Chinese Edition

16007721

USB cable

Instrument Guide | 45

Appendix A Bio-Rad’s Real-Time PCR Detection Systems and CFX Maestro Software Catalog Numbers

46 | CFX Duet Real-Time PCR System

Appendix B Recommended Plastic Consumables
Plastic Consumables for the CFX Duet System
The CFX Duet instrument accepts low-profile 0.2 ml plates and tubes. Bio-Rad recommends HSP9655— Hard-Shell low-profile 96-well skirted PCR plates with white shell and white wells for optimal
results.
The following additional plastic consumables will fit the system but may need to be validated andoptimized for specific workflows:n

MLL9601 — Multiplate low-profile 96-well unskirted PCR plates with clear wellsn

MLL9651 — Multiplate low-profile 96-well unskirted PCR plates with white wellsn

HSP9601 — Hard-Shell low-profile 96-well skirted PCR plates with white shell and clear wellsn

TLS0801 — Low-profile 0.2 ml 8-tube PCR strips without caps, clearn

TLS0851 — Low-profile 0.2 ml 8-tube PCR strips without caps, whiten

TCS0803 — Optical flat 8-cap strip, for 0.2 ml PCR tubes and plates

Plate Seals and Plate Sealer
For optimal results, Bio-Rad recommends the following plate seals:n

MSB1001 — Microseal 'B' adhesive seals, optically clear (strong adhesive-based)n

MSC1001 — Microseal 'C' optical seals, optically clear (pressure-activated, adhesive-based)

Instrument Guide | 47

Appendix B Recommended Plastic Consumables

48 | CFX Duet Real-Time PCR System

Appendix C Maintenance and TroubleshootingThis appendix explains how to clean and maintain the CFX Duet Real-Time PCR Detection Systemand how to troubleshoot problems that you might have. If you need to return the system to Bio-Rad,
see Returning the CFX Duet Real-Time PCR Detection System to Bio-Rad.

Cleaning and Maintaining the CFX Duet Real-Time PCR
Detection System
The CFX Duet instrument requires little maintenance for proper operation and precise thermal control.
However, with long and constant use, the instrument will require some cleaning and othermaintenance.
CFX Duet instruments include a sensitive optical shuttle system that moves quickly during datacollection and a sample block that must heat and cool quickly. Contamination of these components
can interfere with thermal cycling and data collection.
Never allow a reaction to run with an open or leaking sample lid. The reagents could escape and coat
the block, inner lid, and optical head in the shuttle system. Excessive dirt can dim the signal, andfluorescence contamination can create excessive background signal. The shuttle system cannot becleaned except by trained Bio-Rad service engineers.
Avoid contaminating your CFX Duet instrument by following these guidelines:n

Always clean the outside of any containers before placing them in the block.

n

Never run a reaction with a seal that is open, loose, punctured, or otherwise damaged because
you could contaminate the block, inner lid, and optical system.

nNever run a PCR or real-time PCR reaction with volatile reagents that could explode andcontaminate the block, inner lid, and optical system.

nNever run a real-time PCR reaction with volatile reagents that could rupture the sample container.

n

Clean the reaction block and inner lid periodically to prevent the buildup of dirt, biohazardous
material, or fluorescent solutions (see Table 11 on page 52).
Important: Never clean or otherwise touch the optical system behind the heater plate holes
in the inner lid.

Instrument Guide | 49

Appendix C Maintenance and Troubleshooting

50 | CFX Duet Real-Time PCR System

Cleaning and Maintaining the CFX Duet Real-Time PCR Detection SystemnClean the outer surface of the CFX Duet instrument on a regular schedule to remove any debris
or dirt that might interfere with proper function (see Table 11 on page 52). Clean the instrument toprevent damage to the air intake or reaction bay.

Important: For instructions on handling and cleaning radioactive or biohazardous materials,
consult the guidelines for radiation safety and biosafety provided by your institution. These
guidelines also include hazardous materials disposal methods.

Instrument Guide | 51

Appendix C Maintenance and Troubleshooting

CFX Duet Real-Time PCR Detection System Cleaning and Maintenance Safety
Warnings
When cleaning and maintaining the CFX Duet instrument, always consider and follow the warnings
listed in Table 11 that follows.
Table 11. Cleaning and maintenance safety warnings

Warning
To prevent electrical shock, always turn off and unplug the instrument from theelectrical outlet before cleaning it.

A thermal cycler operates at temperatures high enough to cause serious burns. Always
allow the entire instrument to return to room temperature before cleaning.

When handling biohazardous or radioactive samples, adhere to the recommended
precautions and guidelines specific to your laboratory and location. These guidelines
should include cleaning, monitoring, and disposal methods for the hazardous material(s) you are using.
In addition, as identified above, there is a small risk of explosion, or of expulsion ofliquids or vapors from the sample container(s). When working with hazardous materials
the risk of injury from expelled material is compounded with the risk that the hazardous
material themselves could be dispersed in and around the instrument. Users shouldtake appropriate precautions for such a situation.

52 | CFX Duet Real-Time PCR System

Cleaning and Maintaining the CFX Duet Real-Time PCR Detection System

CFX Duet Real-Time PCR Detection System Maintenance
Table 12 lists the components of the CFX Duet instrument that require maintenance.
Table 12. CFX Duet instrument maintenance

Component

ActionAir vents

Use a soft brush, damp cloth, or vacuum cleaner to remove light dust
from the air vents. Remove any heavy dust that is deep in the ventswith a vacuum cleaner.

Tip: Cleaning the vents allows sufficient airflow for precise
thermal control during a run.
Outside case of the system

Use a damp cloth or tissue to clean spills off the outside case. If
needed, use a mild soap solution and remove the residue completely.

Tip: Cleaning the outside case prevents corrosion.
Sample block

Important: Clean spills immediately to prevent them from drying
inside wells.
Use disposable plastic pipettes with water (recommended), 95%
ethanol, or a 1:100 dilution of bleach in water. Always rinse the wellswith water several times to remove all traces of ethanol, bleach, orsoap.

Note: Consider the following when cleaning the sample block:n If left in the block wells, bleach, ethanol, or soap could corrode theblock and/or destroy tubes and microplates during a run. Always
rinse the block thoroughly after cleaning it with any solution otherthan water.

n Never clean the sample block with strong alkaline solutions (strong
soap, ammonia, or highly concentrated bleach). Never use corrosiveor abrasive cleaning solutions. These cleaning agents can damage
the block, preventing precise thermal control.

n Never heat the block with cleaning solution on or in it. Heating theblock with cleaning solution damages the block and lid, and cancontaminate the optics.
If oil is used, the wells must be cleaned thoroughly and often. Use of oilin the wells is not recommended. Use a solution of 95% ethanol toclean oil on the sample block. Do not allow oil to build up in the block.

Instrument Guide | 53

Appendix C Maintenance and Troubleshooting

Table 12. CFX Duet instrument maintenance, continued

Component

ActionInner lid of the sample blockUse a soft cloth and water to remove debris and solutions from theinner lid surface.

Tip: Cleaning the inner lid improves precise sample heating andcooling.

Important: Never use abrasive detergents or rough materials thatscratch the surface.

54 | CFX Duet Real-Time PCR System

Maintaining Sufficient Air Flow

Maintaining Sufficient Air Flow
The CFX Duet instrument requires sufficient air flow to heat and cool precisely to the correct target
temperature. If the flow of air is blocked, the thermal cycler cannot ramp to the correct temperature in thespecified time. This section explains how to test the air flow and how to fix low or warm air flow.

Testing for Sufficient Air Flow
The air flow is sufficient when the instrument heats and cools to the correct target temperatures
promptly. Bio-Rad suggests that you test the air flow when you set up the CFX Duet instrument in a
new location. You can also measure the air temperature at any time to ensure sufficient air flow.

To determine the presence of sufficient air flow
1.

Set up and start the instrument.

2.

Adjust the local environment for typical conditions:

3.

nTurn on any nearby equipment, such as fans or other instruments.

nOpen any window blinds to reproduce typical conditions during a run.

Run a typical PCR protocol for 30 min.
If more than one instrument is in the area, run a protocol on all instruments at the same time.
Note: Samples are not required for the test runs. However, you must include an empty
microplate or capped strip tubes. The lid does not heat correctly if it touches the sample
block.

4.

Measure the air temperature at the air intake vents of the instrument.
If the air intake temperature increases above 31°C, see the following section Fixing Insufficient Air
Flow.

Fixing Insufficient Air Flow
If the air temperature near the instrument is above 31°C, make one or more of the following changes
to increase the flow of cooler air around the instrument:nAdjust air conditioning to lower the ambient air temperature.

nMove the instrument to another location.

nProvide more space around the instrument and between adjacent instruments. Arrange
instruments so that the warm exhaust air from one instrument does not enter the air intake vents ofanother.

Instrument Guide | 55

Appendix C Maintenance and TroubleshootingnShield the instrument from heat sources such as radiators, heat-producing instruments, andbright sunlight.

Replacing Fuses
Fuses on the CFX Duet instrument are designed to blow in case of severe power surges or othercauses of electrical short. This protects both the user and the instrument from excessive, potentiallydamaging electric currents. Fuses on the CFX Duet instrument rarely need to be replaced. However,some institutions prefer to replace fuses on a regular basis to maintain uninterrupted operation.
If the instrument does not turn on, first verify that the power cord is plugged in to a functioning powersource. Also, verify that the power cord and power source are within the specifications for this
instrument.
Important: Do not attempt to replace the power cord on the CFX Duet instrument. Instead, contact
Bio-Rad Technical Support.
Finally, verify that the fuses are intact. If the fuses are broken or burned, replace the fuses. This section
explains how to view and replace fuses on the CFX Duet instrument.
Tip: The instrument utilizes two 10 A, 250 V, 5 x 20 mm, fast blow fuses.
WARNING! To prevent electrical shock, always turn off and unplug the instrumentfrom the electrical outlet before checking the fuses.

To view and replace the fuses
1.

Verify that you have turned off the instrument and unplugged the power cord from the back of the
instrument.
Important: You must unplug the power cord from the instrument to open the fuse door. Trying
to open the fuse door while the cord is plugged in could damage the door.

2.

Using your finger tip, gently pull the black fuse door on the back of the instrument out toward you.

56 | CFX Duet Real-Time PCR System

Replacing Fuses

3.

Using a small flathead screw driver, gently pry out the red fuse holder until you can grasp it withyour fingers.

4.

When you can firmly grasp the fuse holder, pull it straight out of the instrument.

5.

Using a finger tip, gently pry the fuse out of the fuse holder.

6.

The fuse holder contains two fuses, one on each side. You must inspect both fuses.

Instrument Guide | 57

Appendix C Maintenance and Troubleshooting

A bad fuse displays a break or burned spot in the internal metal filament, or measures open with
an Ohm meter. A good fuse has an internal metal filament or measures short (< 1 Ohm). If a fuseis bad or damaged, replace it with a new fuse of the same type and rating.
Note: Some fuses used in the CFX Duet instrument are made from ceramic and cannot be
visually inspected. In this case, you must use an Ohm meter to determine whether the fuse isgood. Alternatively, you can replace the fuse with a known good one without checking.

7.

Assuming the prongs indicate the front of the fuse holder, insert the back end of a 10 A, 250 V, 5 x
20 mm, fast blow fuse into the middle bracket. Ensure the front end of the fuse faces the prongs.
For example:

8.

With the flat red square on the fuse holder facing upward, insert the fuse holder into the
instrument and firmly press it into position.

58 | CFX Duet Real-Time PCR System

Replacing Fuses

9.

Close the fuse door, insert the power cord, and turn on the instrument.
Caution: If the CFX Duet instrument repeatedly blows one or more fuses, there could
be an internal problem with the instrument. Contact Bio-Rad Technical Support forassistance in determining whether it is safe to replace the fuses again or if the
instrument should be repaired.

Instrument Guide | 59

Appendix C Maintenance and Troubleshooting

Upgrading Software and Firmware on CFX Duet Real-Time
PCR Systems
You can only upgrade CFX Duet instrument firmware and CFX Maestro software in CFX Maestro.
Note: Depending on the type of upgrade, this process can take several minutes to complete.

Updating CFX Maestro Software and CFX Duet Real-Time PCR Detection System
Firmware
When connected to the internet, CFX Maestro automatically checks for both CFX Maestro and CFX
Duet updates each time it starts. CFX Maestro can automatically perform the CFX Duet update.
When updates are available, the Software and Instruments Updates dialog box automatically appears.

Important: CFX Duet instruments requiring an update appear with a red alert. You must performthe required update in order to run experiments on that instrument.
When both software and system updates are performed, the software is updated first. In order todownload the latest updates, the computer running CFX Maestro instrument must be connected to theinternet. Close all data analysis windows and ensure that all instruments are idle before beginning the
update.
When the software update completes, CFX Maestro restarts and then begins the CFX Duet updates.
CFX Maestro determines whether the instrument software is compatible with the version of CFX
Maestro that is installed. If the instrument software is not compatible, this update begins automatically.
No internet connection is required for this operation.
Note: You must restart the instrument after the update completes.

60 | CFX Duet Real-Time PCR System

Shutting Down the CFX Duet Real-Time PCR Detection System

To perform software and system updatesu

In the Software and Instrument Updates dialog box, do one of the following:n

If either a software or system update is available, select the appropriate checkbox and click
Update.

n

If both software and instrument updates are available, select both checkboxes and click
Update.
Tip: In this case, updating the software requires an instrument update for any connected
instruments to ensure proper communication.

To hide the Software and Instrument Updates dialog box alertu

To hide the Updates dialog box when new updates are available, clear the Automatically show
this window when updates are available checkbox.
Important: If an attached instrument requires an update, the Software and Instrument
Updates dialog box automatically appears even if this checkbox is cleared. You must performthe requisite update in order to run experiments on that instrument.
To display the Updates dialog box, on the Home window select Help > Check for Updates andselect the Notify when updates are available checkbox.

Shutting Down the CFX Duet Real-Time PCR Detection
SystemFollow these instructions to safely and completely shut down the CFX Duet instrument.

To shut down the CFX Duet instrument
1.

Ensure that no protocol is running and the instrument is no longer in use.

2.

In the CFX Maestro Detected Instruments pane, select the CFX Duet instrument.

3.

Click Open Lid.

4.

Remove the sample plate located on the block of the CFX Duet instrument.

5.

Click Close Lid.

6.

Press the power switch on the back panel of the instrument to power down the system.

Instrument Guide | 61

Appendix C Maintenance and Troubleshooting

Returning the CFX Duet Real-Time PCR Detection System to
Bio-RadImportant: Should you need to return your CFX Duet instrument to Bio-Rad, your Bio-Rad
Technical Support specialist will provide instructions for decontaminating, packaging, andshipping the instrument. Before you return the instrument, you must install the shipping plate andshipping screw. This section explains these tasks.
Note: Locate the shipping screw and shipping plate that you saved when you installed thesystem. You will need these items to properly package the system. Bio-Rad will send you thepackaging material required to safely return the system.
Important: Before you begin, ensure that you have completely backed up all data files to ashared network drive or a USB drive.

Installing the Shipping Plate and Screw
To install the shipping plate and screw
1.

Start the CFX Duet instrument and log in to CFX Maestro.

2.

In CFX Maestro, transfer all data files from the CFX Duet instrument to a computer running CFX
Maestro. For instructions on how to transfer user data, refer to the CFX Maestro User Guide.

3.

To install the shipping screw:a.

Right-click on the instrument in the Detected Instruments pane of CFX Maestro and selectProperties.

b.

Click the Shipping Screw tab.

c.

Click the Open Lid button.

d.

Place the shipping plate on the CFX Duet instrument sample block.

e.

Click the Close Lid button.

f.

Click the Lock Lid button.

g.

Confirm that the CFX Duet instrument shipping plate is installed, and click the Yes button in
the confirmation dialog.

62 | CFX Duet Real-Time PCR System

Returning the CFX Duet Real-Time PCR Detection System to Bio-Rad

4.

Remove the beige plug (store it in a safe place) and then install the shipping screw, turning it
clockwise until it is securely in place.

5.

Click the Screw Installed button to confirm that the shipping screw is installed.

6.

Log out of CFX Maestro and then shut down the CFX Duet instrument.

7.

Package and ship the system to Bio-Rad according to the instructions provided by Bio-Rad.

Instrument Guide | 63

Appendix C Maintenance and Troubleshooting

64 | CFX Duet Real-Time PCR System

Appendix D Bio-Rad Free and Open-Source
Notices for PCR ProductsThis document includes licensing information relating to free, open- source, and public- source
software and data (together, the “MATERIALS”) included with or used to develop Bio-Rad products
and services. The terms of the applicable free, open-source, and public-source licenses (each an“OPEN LICENSE”) govern Bio-Rad’s distribution and your use of the MATERIALS. Bio-Rad and thethird-party authors, licensors, and distributors of the MATERIALS disclaim all warranties and liability
arising from all use and distribution of the MATERIALS. To the extent the OSS is provided under anagreement with Bio-Rad that differs from the applicable OSS LICENSE, those terms are offered by
Bio-Rad alone.
Bio- Rad has reproduced below copyright and other licensing notices appearing within the
MATERIALS. While Bio- Rad seeks to provide complete and accurate copyright and licensing
information for all MATERIALS, Bio-Rad does not represent or warrant that the following information iscomplete, correct, or error-free. MATERIALS recipients are encouraged to (a) investigate the identified
MATERIALS to confirm the accuracy of the licensing information provided and (b) notify Bio-Rad of
any inaccuracies or errors found in this document so that Bio- Rad may update this document
accordingly.
Certain OPEN LICENSES (such as the Affero General Public Licenses, Common Development andDistribution Licenses, Common Public License, Creative Commons Share- Alike License, Eclipse
Public License, Mozilla Public Licenses, GNU General Public Licenses, GNU Library/Lesser General
Public Licenses, and Open Data Commons Open Database License) require that the source materials
be made available to recipients or other requestors under the terms of the same OPEN LICENSE.
The corresponding open source software is available for download from the links in the section that
follows.

Instrument Guide | 65

Appendix D Bio-Rad Free and Open-Source Notices for PCR Products

Software Notices
M2Mqtt (paho.mqtt.m2mqtt)
Project homepage/download sites:
https://m2mqtt.wordpress.com/
https://www.nuget.org/packages/M2Mqtt/
Bio-Rad source code site:
https://github.com/bio-rad-lsg-open-source/Mqtt-4.3.0.0
External source code site:
https://github.com/eclipse/paho.mqtt.m2mqtt
Project licensing notices:

Eclipse Foundation Software User Agreement
February 1, 2011
Usage Of Content
THE ECLIPSE FOUNDATION MAKES AVAILABLE SOFTWARE, DOCUMENTATION,
INFORMATION AND/OR OTHER MATERIALS FOR OPEN SOURCE PROJECTS(COLLECTIVELY "CONTENT"). USE OF THE CONTENT IS GOVERNED BY THE TERMS
AND CONDITIONS OF THIS AGREEMENT AND/OR THE TERMS AND CONDITIONS OF
LICENSE AGREEMENTS OR NOTICES INDICATED OR REFERENCED BELOW. BY USINGTHE CONTENT, YOU AGREE THAT YOUR USE OF THE CONTENT IS GOVERNED BY
THIS AGREEMENT AND/OR THE TERMS AND CONDITIONS OF ANY APPLICABLE
LICENSE AGREEMENTS OR NOTICES INDICATED OR REFERENCED BELOW. IF YOU
DO NOT AGREE TO THE TERMS AND CONDITIONS OF THIS AGREEMENT AND THE
TERMS AND CONDITIONS OF ANY APPLICABLE LICENSE AGREEMENTS OR NOTICESINDICATED OR REFERENCED BELOW, THEN YOU MAY NOT USE THE CONTENT.
Applicable Licenses
Unless otherwise indicated, all Content made available by the Eclipse
Foundation is provided to you under the terms and conditions of the
Eclipse Public License Version 1.0 ("EPL"). A copy of the EPL isprovided with this Content and is also available at
http://www.eclipse.org/legal/epl-v10.html. For purposes of the EPL,
"Program" will mean the Content.

66 | CFX Duet Real-Time PCR System

Software Notices

Content includes, but is not limited to, source code, object code,
documentation and other files maintained in the Eclipse Foundation
source code repository ("Repository") in software modules ("Modules")and made available as downloadable archives ("Downloads").
l

l

l

l

Content may be structured and packaged into modules tofacilitate delivering, extending, and upgrading the Content.
Typical modules may include plug-ins ("Plug-ins"), plug-in
fragments ("Fragments"), and features ("Features").
Each Plug-in or Fragment may be packaged as a sub-directory or
JAR (Java™ ARchive) in a directory named "plugins".
A Feature is a bundle of one or more Plug-ins and/or Fragments
and associated material. Each Feature may be packaged as a subdirectory in a directory named "features". Within a Feature,
files named "feature.xml" may contain a list of the names andversion numbers of the Plug-ins and/or Fragments associated withthat Feature.
Features may also include other Features ("Included Features").
Within a Feature, files named "feature.xml" may contain a listof the names and version numbers of Included Features.

The terms and conditions governing Plug-ins and Fragments should becontained in files named "about.html" ("Abouts"). The terms andconditions governing Features and Included Features should becontained in files named "license.html" ("Feature Licenses"). Abouts
and Feature Licenses may be located in any directory of a Download or
Module including, but not limited to the following locations:
l
l
l
l
l

The top-level (root) directoryPlug-in and Fragment directories
Inside Plug-ins and Fragments packaged as JARs
Sub-directories of the directory named "src" of certain Plug-ins
Feature directories

Note: if a Feature made available by the Eclipse Foundation is
installed using the Provisioning Technology (as defined below), youmust agree to a license ("Feature Update License") during the
installation process. If the Feature contains Included Features, theFeature Update License should either provide you with the terms andconditions governing the Included Features or inform you where you
can locate them. Feature Update Licenses may be found in the"license" property of files named "feature.properties" found within a
Feature. Such Abouts, Feature Licenses, and Feature Update Licenses
contain the terms and conditions (or references to such terms andInstrument Guide | 67

Appendix D Bio-Rad Free and Open-Source Notices for PCR Productsconditions) that govern your use of the associated Content in that
directory.
THE ABOUTS, FEATURE LICENSES, AND FEATURE UPDATE LICENSES MAY REFER
TO THE EPL OR OTHER LICENSE AGREEMENTS, NOTICES OR TERMS AND
CONDITIONS. SOME OF THESE OTHER LICENSE AGREEMENTS MAY INCLUDE (BUT
ARE NOT LIMITED TO):
l

l

l

l

l

l

Eclipse Distribution License Version 1.0 (available at
http://www.eclipse.org/licenses/edl-v1.0.html)
Common Public License Version 1.0 (available at
http://www.eclipse.org/legal/cpl-v10.html)
Apache Software License 1.1 (available at
http://www.apache.org/licenses/LICENSE)
Apache Software License 2.0 (available at
http://www.apache.org/licenses/LICENSE-2.0)
Metro Link Public License 1.00 (available at
http://www.opengroup.org/openmotif/supporters/metrolink/license.html)
Mozilla Public License Version 1.1 (available at
http://www.mozilla.org/MPL/MPL-1.1.html)

IT IS YOUR OBLIGATION TO READ AND ACCEPT ALL SUCH TERMS AND
CONDITIONS PRIOR TO USE OF THE CONTENT. If no About, Feature License,or Feature Update License is provided, please contact the Eclipse
Foundation to determine what terms and conditions govern thatparticular Content.

Use of Provisioning Technology
The Eclipse Foundation makes available provisioning software,examples of which include, but are not limited to, p2 and the Eclipse
Update Manager ("Provisioning Technology") for the purpose ofallowing users to install software, documentation, information and/or
other materials (collectively "Installable Software"). This
capability is provided with the intent of allowing such users to
install, extend and update Eclipse-based products. Information about
packaging Installable Software is available at
http://eclipse.org/equinox/p2/repository_packaging.html("Specification").
You may use Provisioning Technology to allow other parties to install
Installable Software. You shall be responsible for enabling theapplicable license agreements relating to the Installable Software to
be presented to, and accepted by, the users of the Provisioning

68 | CFX Duet Real-Time PCR System

Software Notices

Technology in accordance with the Specification. By using
Provisioning Technology in such a manner and making it available inaccordance with the Specification, you further acknowledge youragreement to, and the acquisition of all necessary rights to permit
the following:
1. A series of actions may occur ("Provisioning Process") in which auser may execute the Provisioning Technology on a machine ("Target
Machine") with the intent of installing, extending or updating thefunctionality of an Eclipse-based product.
2. During the Provisioning Process, the Provisioning Technology may
cause third party Installable Software or a portion thereof to beaccessed and copied to the Target Machine.
3. Pursuant to the Specification, you will provide to the user theterms and conditions that govern the use of the Installable Software("Installable Software Agreement") and such Installable Software
Agreement shall be accessed from the Target Machine in accordancewith the Specification. Such Installable Software Agreement must
inform the user of the terms and conditions that govern the
Installable Software and must solicit acceptance by the end user inthe manner prescribed in such Installable Software Agreement. Uponsuch indication of agreement by the user, the provisioning Technology
will complete installation of the Installable Software.

Cryptography
Content may contain encryption software. The country in which you are
currently may have restrictions on the import, possession, and use,
and/or re-export to another country, of encryption software. BEFORE
using any encryption software, please check the country's laws,
regulations and policies concerning the import, possession, or use,
and re-export of encryption software, to see if this is permitted.
Java and all Java-based trademarks are trademarks of Oracle
Corporation in the United States, other countries, or both.
/LICENSE:
See EPL-1.0 in the Standard OSS License Text appendix to this document.

Instrument Guide | 69

Appendix D Bio-Rad Free and Open-Source Notices for PCR ProductsStandard Open License Text
EPL 1.0
Eclipse Public License - v 1.0
THE ACCOMPANYING PROGRAM IS PROVIDED UNDER THE TERMS OF THIS ECLIPSE
PUBLIC LICENSE ("AGREEMENT"). ANY USE, REPRODUCTION OR DISTRIBUTION
OF THE PROGRAM CONSTITUTES RECIPIENT'S ACCEPTANCE OF THIS AGREEMENT.
1. DEFINITIONS"Contribution" means:a) in the case of the initial Contributor, the initial code anddocumentation distributed under this Agreement, andb) in the case of each subsequent Contributor:i) changes to the Program, andii) additions to the Program;where such changes and/or additions to the Program originate from andare distributed by that particular Contributor. A Contribution'originates' from a Contributor if it was added to the Program bysuch Contributor itself or anyone acting on such Contributor'sbehalf. Contributions do not include additions to the Program which:(i) are separate modules of software distributed in conjunction with
the Program under their own license agreement, and (ii) are notderivative works of the Program.
"Contributor"
Program.

means

any

personorentitythat

distributes

the"Licensed Patents" mean patent claims licensable by a Contributor
which are necessarily infringed by the use or sale of its
Contribution alone or when combined with the Program.
"Program" means the Contributions distributed in accordance with thisAgreement.
"Recipient" means anyone who receives
Agreement, including all Contributors.

70 | CFX Duet Real-Time PCR Systemthe

Program

under

this

Standard Open License Text

2. GRANT OF RIGHTSa) Subject to the terms of this Agreement, each Contributor hereby
grants Recipient a non-exclusive, worldwide, royalty-free copyrightlicense to reproduce, prepare derivative works of, publicly display,
publicly perform, distribute and sublicense the Contribution of such
Contributor, if any, and such derivative works, in source code andobject code form.
b) Subject to the terms of this Agreement, each Contributor hereby
grants Recipient a non- exclusive, worldwide, royalty- free patent
license under Licensed Patents to make, use, sell, offer to sell,
import and otherwise transfer the Contribution of such Contributor,
if any, in source code and object code form. This patent license
shall apply to the combination of the Contribution and the Program
if, at the time the Contribution is added by the Contributor, such
addition of the Contribution causes such combination to be covered by
the Licensed Patents. The patent license shall not apply to any othercombinations which include the Contribution. No hardware per se islicensed hereunder.
c) Recipient understands that although each Contributor grants thelicenses to its Contributions set forth herein, no assurances areprovided by any Contributor that the Program does not infringe thepatent or other intellectual property rights of any other entity.
Each Contributor disclaims any liability to Recipient for claims
brought by any other entity based on infringement of intellectual
property rights or otherwise. As a condition to exercising the rights
and licenses granted hereunder, each Recipient hereby assumes sole
responsibility to secure any other intellectual property rights
needed, if any. For example, if a third party patent license isrequired to allow Recipient to distribute the Program, it isRecipient's
responsibility
toacquirethat
licensebeforedistributing the Program.
d) Each Contributor represents that to its knowledge it has
sufficient copyright rights in its Contribution, if any, to grant the
copyright license set forth in this Agreement.
3. REQUIREMENTS
A Contributor may choose to distribute the Program in object code
form under its own license agreement, provided that:a) it complies with the terms and conditions of this Agreement; andInstrument Guide | 71

Appendix D Bio-Rad Free and Open-Source Notices for PCR Productsb) its license agreement:i) effectively disclaims on behalf of all Contributors all warranties
and conditions, express and implied, including warranties orconditions of title and non-infringement, and implied warranties orconditions of merchantability and fitness for a particular purpose;ii) effectively excludes on behalf of all Contributors all liability
for damages, including direct, indirect, special, incidental andconsequential damages, such as lost profits;iii) states that any provisions which differ from this Agreement are
offered by that Contributor alone and not by any other party; andiv) states that source code for the Program is available from such
Contributor, and informs licensees how to obtain it in a reasonable
manner on or through a medium customarily used for software exchange.
When the Program is made available in source code form:a) it must be made available under this Agreement; andb) a copy of this Agreement must be included with each copy of theProgram.
Contributors may not remove or alter any copyright notices contained
within the Program.
Each Contributor must identify itself as the originator of its
Contribution, if any, in a manner that reasonably allows subsequent
Recipients to identify the originator of the Contribution.
4. COMMERCIAL DISTRIBUTION
Commercial
distributorsofsoftwaremay
acceptcertain
responsibilities with respect to end users, business partners and thelike. While this license is intended to facilitate the commercial useof the Program, the Contributor who includes the Program in acommercial product offering should do so in a manner which does notcreate potential liability for other Contributors. Therefore, if aContributor includes the Program in a commercial product offering,such Contributor ("Commercial Contributor") hereby agrees to defend
and indemnify every other Contributor ("Indemnified Contributor")
against any losses, damages and costs (collectively "Losses") arisingfrom claims, lawsuits and other legal actions brought by a third
party against the Indemnified Contributor to the extent caused by theacts or omissions of such Commercial Contributor in connection with72 | CFX Duet Real-Time PCR System

Standard Open License Text

its distribution of the Program in a commercial product offering. The
obligations in this section do not apply to any claims or Losses
relating to any actual or alleged intellectual property infringement.
In order to qualify, an Indemnified Contributor must: a) promptly
notify the Commercial Contributor in writing of such claim, and b)
allow the Commercial Contributor to control, and cooperate with theCommercial Contributor in, the defense and any related settlement
negotiations. The Indemnified Contributor may participate in any suchclaim at its own expense.
For example, a Contributor might include the Program in a commercial
product offering, Product X. That Contributor is then a Commercial
Contributor. If that Commercial Contributor then makes performance
claims, or offers warranties related to Product X, those performance
claimsandwarranties
aresuchCommercial
Contributor's
responsibility alone. Under this section, the Commercial Contributor
would have to defend claims against the other Contributors related tothose performance claims and warranties, and if a court requires anyother Contributor to pay any damages as a result, the Commercial
Contributor must pay those damages.
5. NO WARRANTY
EXCEPT AS EXPRESSLY SET FORTH IN THIS AGREEMENT, THE PROGRAM ISPROVIDED ON AN "AS IS" BASIS, WITHOUT WARRANTIES OR CONDITIONS OF ANY
KIND, EITHER EXPRESS OR IMPLIED INCLUDING, WITHOUT LIMITATION, ANY
WARRANTIES OR CONDITIONS OF TITLE, NON-INFRINGEMENT, MERCHANTABILITY
OR FITNESS FOR A PARTICULAR PURPOSE. Each Recipient is solely
responsible for determining the appropriateness of using anddistributing the Program and assumes all risks associated with its
exercise of rights under this Agreement, including but not limited to
the risks and costs of program errors, compliance with applicablelaws, damage to or loss of data, programs or equipment, andunavailability or interruption of operations.
6. DISCLAIMER OF LIABILITY
EXCEPT AS EXPRESSLY SET FORTH IN THIS AGREEMENT, NEITHER RECIPIENTNOR ANY CONTRIBUTORS SHALL HAVE ANY LIABILITY FOR ANY DIRECT,INDIRECT, INCIDENTAL, SPECIAL, EXEMPLARY, OR CONSEQUENTIAL DAMAGES(INCLUDING WITHOUT LIMITATION LOST PROFITS), HOWEVER CAUSED AND ON
ANY THEORY OF LIABILITY, WHETHER IN CONTRACT, STRICT LIABILITY, OR
TORT (INCLUDING NEGLIGENCE OR OTHERWISE) ARISING IN ANY WAY OUT OF
THE USE OR DISTRIBUTION OF THE PROGRAM OR THE EXERCISE OF ANY RIGHTS

Instrument Guide | 73

Appendix D Bio-Rad Free and Open-Source Notices for PCR Products

GRANTED HEREUNDER,DAMAGES.

EVENIF

ADVISED

OF

THE

POSSIBILITY

OF

SUCH

7. GENERAL
If any provision of this Agreement is invalid or unenforceable underapplicable law, it shall not affect the validity or enforceability of
the remainder of the terms of this Agreement, and without further
action by the parties hereto, such provision shall be reformed to theminimum
extentnecessary
to
make
such
provisionvalid
andenforceable.
If Recipient institutes patent litigation against any entity(including a cross-claim or counterclaim in a lawsuit) alleging that
the Program itself (excluding combinations of the Program with othersoftware or hardware) infringes such Recipient's patent(s), then such
Recipient's rights granted under Section 2(b) shall terminate as ofthe date such litigation is filed.
All Recipient's rights under this Agreement shall terminate if it
fails to comply with any of the material terms or conditions of thisAgreement and does not cure such failure in a reasonable period oftime after becoming aware of such noncompliance. If all Recipient's
rights under this Agreement terminate, Recipient agrees to cease use
and distribution of the Program as soon as reasonably practicable.
However, Recipient's obligations under this Agreement and any
licenses granted by Recipient relating to the Program shall continue
and survive.
Everyone is permitted to copy and distribute copies of thisAgreement, but in order to avoid inconsistency the Agreement is
copyrighted and may only be modified in the following manner. The
Agreement Steward reserves the right to publish new versions(including revisions) of this Agreement from time to time. No one
other than the Agreement Steward has the right to modify thisAgreement. The Eclipse Foundation is the initial Agreement Steward.
The Eclipse Foundation may assign the responsibility to serve as theAgreement Steward to a suitable separate entity. Each new version of
the Agreement will be given a distinguishing version number. The
Program (including Contributions) may always be distributed subject
to the version of the Agreement under which it was received. In
addition, after a new version of the Agreement is published,
Contributor may elect to distribute the Program (including its
Contributions) under the new version. Except as expressly stated in
Sections 2 (a) and 2 (b) above, Recipient receives no rights or

74 | CFX Duet Real-Time PCR System

Standard Open License Text

licenses to the intellectual property of any Contributor under thisAgreement, whether expressly, by implication, estoppel or otherwise.
All rights in the Program not expressly granted under this Agreement
are reserved.
This Agreement is governed by the laws of the State of New York andthe intellectual property laws of the United States of America. No
party to this Agreement will bring a legal action under thisAgreement more than one year after the cause of action arose. Eachparty waives its rights to a jury trial in any resulting litigation.

Instrument Guide | 75

Appendix D Bio-Rad Free and Open-Source Notices for PCR Products

76 | CFX Duet Real-Time PCR System

Appendix E References1.

Breslauer KJ et al. (1986). Predicting DNA duplex stability from the base sequence. Proc Natl
Acad Sci USA 83, 3,746–3,750.

2.

Sugimoto N et al. (1996). Improved thermodynamic parameters and helix initiation factor to
predict stability of DNA duplexes. Nucleic Acids Res 24, 4,501–4,505.

Instrument Guide | 77Appendix E References78 | CFX Duet Real-Time PCR System

Bio-RadLaboratories, Inc.
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