Real-Time PCR

CFX Duet
Real-Time PCR System

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

CFX DUET REAL-TIME PCR SYSTEM

POWERFUL YET ACCESSIBLE

GENOMICS ANALYSIS

With the CFX Duet Real-Time PCR System, Bio-Rad™ Laboratories, Inc. makes powerfuland consistent quantitative PCR (qPCR) accessible to every laboratory. High-end thermal
performance and the proprietary, accurate optical shuttle system from our CFX Opus
Systems now comes in an easy-to-use, dual color format. Our popular CFX Maestro
Software drives this system and makes essential genomics analyses approachable forthe economical lab.
Search your mobile app store for CFX qPCR Systems to see, using augmented reality,
how the system fits into your lab, or visit bio-rad.com/CFXDuet for links and details.

2 | BIO-RAD LABORATORIES, INC.

HAVE CONFIDENCE IN YOUR ENTIRE GENOMICS WORKFLOW
Unlike many suppliers, Bio-Rad offers everything you need for real-time PCR experiments, from sample
preparation to data analysis. Simplify and optimize your workflow with our robust suite of reagents,
instruments, and software, so that you can get publication-quality data every time.

Cell Lysis and RNA Isolation■ Aurum Total RNA Mini Kit■ Aurum Total RNA 96 Kit■ Aurum Total RNA Fatty and Fibrous Tissue Kit■ PureZOL RNA Isolation Reagent■ SingleShot Cell Lysis Kits

Target Selection■ PrimePCR Assays■ PrimePCR Pathway Panels■ PrimePCR Custom Plates

Reaction Setup■ SsoAdvanced Universal Supermixes■ iTaq Universal Supermixes■ Reliance One-Step Multiplex Supermix■ iTaq One-Step Universal Kits
■ SingleShot Cell Lysis RT-qPCR Kits
■ iScript Reverse Transcription Supermix for RT-qPCR■ iScript gDNA Clear cDNA Synthesis Kit■ PCR Plastics

Detection■ CFX Family of Real-Time PCR SystemsAnalysis■ CFX Maestro Desktop Software

BIO-RAD LABORATORIES, INC. | 3CFX DUET REAL-TIME PCR SYSTEM

ROBUSTOPTICAL DESIGN
Reporter dye:

Channel 1
450–490
FAM

Channel 2515–535
HEX

1.00
0.90
0.80
0.70
0.60
0.50
0.40
0.30
0.20
0.10
0.00
425

450500Detection

550

600
Wavelength, nm

650

700

750

775

650

700

750

775

Channel 1 Channel 2
510–530 560–580
FAM
HEX

Reporter dye:
1.00
0.90
0.80
0.70
0.60
0.50
0.40
0.30
0.200.100.00
425450

500

550

600
Wavelength, nm

Outstanding Duplex Performance
Amplification
5040
RFU (103)

Multiple Data Acquisition Modes
The CFX Duet System can acquire data using several modes.
Choose to acquire data for SYBR® Green I, EvaGreen®, andsingle-color FAM protocols using the fast scan mode or choose toacquire data from both FAM and HEX channels when performing
multiplex protocols. The CFX Duet System includes one channel
with an LED-filter photodiode combination designated for singlecolor fluorescence resonance energy transfer (FRET) experiments,
further expanding your experiment options. FRET mode canexpand your experiment options to applications such as protein
thermal shift (melt) analysis.

Excitation

Normalized absorbanceTwo-Target Multiplexing
The CFX Duet System can discriminate between two targets
in a single reaction well. The optical filter sets are designed tomaximize fluorescence detection for specific dyes in specificchannels. At every position and with every scan, the optics shuttle
is reproducibly centered above each well so the light path is always
fixed and optimal. There is no need to sacrifice data collection in
one of the channels to normalize to a passive reference.

Discrete Channels for Multiplex Data Acquisition

Normalized absorbanceThe solid-state optical technology of the CFX Duet Systemprovides sensitive detection for precise quantification and target
discrimination. Scanning just above the sample plate, the optics
shuttle individually illuminates and detects fluorescence in eachwell with high sensitivity and consistency. This technology differsfrom competing technologies that use a single light source andcamera, eliminating the need for normalization to remove theeffects of varying detection angles. See Bio-Rad bulletin 6047 formore information on our CFX optics.

302010001020246log starting quantity

Cycles304050810Cq

Standard Curve35302520151050As the CFX Duet System’s optical shuttle travels across the plate, light isfocused into the center of each sample well. Side view of the optics shuttle shows
the 450–490 nm LED firing and SYBR® Green emitting at 520 nm into the detector.

Standard
FAM E=97.1% R2=0.999 Slope=–3.394 y-int=39.100
HEX E=94.7% R2=0.999 Slope=–3.455 y-int=39.585

Duplex detection of two plasmids encoding IL-1b and GAPDH.
Both plasmid targets were detected across 10 orders of magnitude(1010 to 10 0 copies) using SsoAdvanced Universal Probes Supermix on
a CFX Duet System. The results demonstrated exceptional performancewith superior efficiency and linearity over a wide dynamic range.
Cq, quantification cycle; RFU, relative fluorescence units.

4 | BIO-RAD LABORATORIES, INC.

CFX DUET REAL-TIME PCR SYSTEMCONSISTENT
THERMAL CYCLING

Large Dynamic Range
The CFX Duet System’s precise thermal performance and optics can achieve
excellent results over 10 logs of input in both FAM and HEX channels.
AAmplification

Standard Curve

605040Cq

RFU (103)

Superior Uniformity
We took a great block and made it even better.
The CFX Duet System uses an improved versionof the CFX Touch Systems’ block to offer
exceptional performance and uniformity while
keeping compatibility with our consumables.
With the best thermal uniformity and accuracy
Bio-Rad has ever produced, you’ll never worry
about using the block from edge to edge again,even with highly sensitive assays.

302010403530252015
105000102030
Cycles4050246log starting quantity

810Standard
FAM E=101.6% R2=0.997 Slope=–3.285 y-int=38.362

BAmplification

Standard Curve

15Cq

RFU (103)20

1054035302520
15105000102030Cycles

4050246log starting quantity

810Standard
HEX E=99.5% R2=0.995 Slope=–3.334 y-int=38.872

Detection of IL-1B in both FAM and HEX channels. A tenfold dilution series (106 to 10 copies) ofplasmid encoding IL-1b was amplified and detected with A, FAM probes and B, HEX probes. Cq,quantification cycle; RFU, relative fluorescence units.
B909075

100Superior thermal uniformity for reproducible results. The temperature measured by probes
in 15 wells across a sample block. A, temperatures across the sample block vary by ±0.3°C for
the CFX Duet System, an improvement of 0.1°C over B, the temperature variability across theCFX Touch Systems’ sample block.
103

102

10* U.S. patent 7,632,464B2.

80Time, sec

100Time, sec

RFU

Gradient Optimization
Not all qPCR systems offer the value of gradientfunctionality. The thermal gradient feature of theCFX Duet System allows you to optimize yourassay in a single experiment, minimizing the useof precious samples and reagents and saving
valuable research time. At any step in a protocol,users can program a temperature gradient of up
to 24°C across the reaction block, with exceptionaltemperature uniformity and reproducibility withineach gradient zone.

ATemperature, °C

The Bio-Rad patented reduced-mass sample block* heats andcools more quickly than standard blocks, improving thermal
uniformity and minimizing edge effects.

01020Cycles

3040

Excellent uniformity. A plasmid template encoding IL-1b diluted to 105 copies/reaction amplified
in the presence of a FAM dye−labeled detection probe with iQ Supermix. Graph shows
96 replicates of 10 μl reactions. Average quantification cycle (Cq) = 19.81 ± 0.10. RFU,relative fluorescence units.

BIO-RAD LABORATORIES, INC. | 5

CFX DUET REAL-TIME PCR SYSTEMEASY-TO-USE

SOFTWARE

Custom data view. View and analyze the data relevant toyour needs on one screen with custom data view.

With CFX Maestro Software You Can:■ 
Perform automatic statistical analysis in seconds — runt-tests or analyze your data using one-way analysis of variance(ANOVA) with just a few mouse clicks■■■

 xtract more meaningful information from each run —Eanalyze data using bar charts, box and whisker plots, dot plots,clustergrams, scatter plots, or volcano plots
 reate and export publication-ready graphics — annotateCgraphs with P values, text, and arrows to call out specific data.
Change colors, fonts, and legends. Export graphs at any sizeand resolution for presentations, posters, or publication
Integrate PrimePCR Assays easily — save time withpredesigned and validated PrimePCR Primers and Plates. Dragand drop your PrimePCR worksheet into CFX Maestro Software
to instantly create your plate setup. Post-run, check run quality
with the PrimePCR controls analysis toolPrecision Melt Analysis Software
Precision Melt Analysis Software imports and analyzes data files
generated by the CFX family of Real-Time PCR Systems to genotype
DNA samples based on their thermal denaturation. The software canbe used for a variety of applications, including gene variant discovery,
single nucleotide polymorphism screening, identification of insertions,deletions, or other unknown mutations, and analysis of methylated
DNA percentage in unknown samples.
The Security You Need
The Security Edition of CFX Maestro Software integrates thepower of the CFX Duet System with tools that enable U.S. FDA
21 CFR Part 11 compliance.
Have confidence in the security of your data, thanks to:
■

■

■

■

■

■

6 | BIO-RAD LABORATORIES, INC.

 andatory password-protected login — valid WindowsM
user profile (local or directory) and password are required ardware protection key (HASP HL key) — key must beHattached to a USB port on the computer to use the softwareFile encryption — files cannot be opened or edited using
other programs
 utomatic file checking — integrity and validity are checkedAeach time a file is opened
 lectronic signatures — more than one electronic signatureE
can be applied to any file that can be opened with the software ime- and date-stamped audit trails — read-only information
Tin the audit trail can be viewed only while the data file is open

CFX DUET REAL-TIME PCR SYSTEMCOMPLEMENTARYPCR PLASTICS AND REAGENTS

Bio-Rad PCR Plastics
Bio-Rad PCR Plastics, specifically our Hard-Shell™ PCR Plates, are the perfect partner for your CFX Duet System.
Hard-Shell PCR Plates offer:
■

■■

■

Automation compatibility
 uperior well-to-well uniformity for reliable and reproducibleS
real-time PCR results
 arp resistance and superior stability for durability duringWrobotic handling, high-speed centrifugation, and storage

The skirt and deck of a
Hard-Shell PCR Plate prevent
warping and shrinkage■ ptional user-readable barcode with alphanumeric labels forOeasy well identification in high-throughput settings
 ootprint and well spacing designed to match American
FNational Standards Institute (ANSI) and Society for Laboratory
Automation and Screening (SLAS) standard dimensions■

Plastics certified free from DNase, RNase, and human DNA■

Choice of 96- or 384-well format

Raised rims allow
for tight sealing

Rigid skirt is ideal
for heat sealingThin-wall polypropylene V-shaped wells enableoptimal thermal transfer and recovery of lowvolume samples

Features of Hard-Shell PCR Plates.

Bio-Rad PCR Reagents
Bio-Rad PCR Reagents offer trusted performance for our Real-Time PCR Systems, including the CFX Duet System.
Our Bio-Rad PCR Reagents include:■■■ dvanced buffer formulations — our PCR reagent buffersA
have been refined to deliver top-notch performance. Use themconfidently with a broad range of complex sample types andtarget regions, as well as under varying reaction parameters(primer concentration, annealing temperature, salt concentration,and cycling protocol)
 niversal reference dyes — a proprietary blend of referenceUdyes means you won’t need different, system-specific dye
concentrations. With our universal reference dyes, reaction
setup is easy and data are less variable. Our reference dyes
are compatible with high ROX, low ROX, and ROX
independent systems
 obust Reliance Reverse Transcriptase — the Reliance
RReverse Transcriptase is genetically engineered for highthermostability and efficient, sensitive cDNA synthesis, evenwith challenging samples and targets■

 owerful Sso7d* fusion DNA polymerase — our Sso7d fusion
PDNA polymerase is designed to succeed where others fail. Itfeatures improved processivity for effective amplification throughregions of strong secondary structures, GC-rich regions, andlonger products, even in the presence of PCR inhibitors

The Best Fit for Your Application
To determine which products will work best for your system, talk to
your local representative, or check out our selector tools:■■

PCR Plastics Selector
PCR and qPCR Reagents Selector* U.S. patent 7,560,260B2.

BIO-RAD LABORATORIES, INC. | 7

Ordering Information
Catalog #Description

Instrument
12016265

CFX Duet Real-Time PCR System, includes power cord, USB cable

SoftwareAll software for Windows unless otherwise noted.
12013758
CFX Maestro Software, includes USB installation drive, video
quick guides, PDF instruction manual
12012832
12013028
12012833
12012834
1845025
Consumables
HSP9655
HSP9601
MSB1001
TLS0851
TLS0801
TCS0803
Reagents
1845098
12010176
172527017250161725095
17005726
12011928

CFX Maestro Software, Security Edition, 1 license
CFX Maestro Software, Security Edition, 5 licenses
CFX Maestro Software, Russian Edition, 1 license
CFX Maestro Software, Chinese Edition, 1 license
Precision Melt Analysis Software, includes 2 user licenses,
installation CD, 2 HASP HL keys, melt calibration kit
Hard-Shell 96-Well PCR Plates, pkg of 50, low profile, thin wall,skirted, white shell/white wells
Hard-Shell 96-Well PCR Plates, pkg of 50, low profile, thin wall,
skirted, white shell/clear wells
Microseal™ 'B' PCR Plate Sealing Film, pkg of 100, optically clear
seals for PCR plates
0.2 ml 8-Tube PCR Strips without Caps, low profile, white
0.2 ml 8-Tube PCR Strips without Caps, low profile, clear0.2 ml Flat PCR Tube 8-Cap Strips, optical, ultraclear
CFX Qualification Plate, 96-well
Reliance One-Step Multiplex RT-qPCR Supermix, 1 ml (1 x 1 ml),
200 x 20 μl reactions
SsoAdvanced Universal SYBR® Green Supermix, 2 ml (2 x 1 ml),
200 x 20 μl reactions
SsoAdvanced Universal Inhibitor-Tolerant SYBR® GreenSupermix, 2 ml (2 x 1 ml), 200 x 20 μl reactions
SingleShot SYBR® Green One-Step Kit for Cell Lysis andRT-qPCR, 100 x 50 μl reactions
SEQuoia Complete Stranded RNA Library Prep Kit, 24 reactions
SEQuoia Dual Indexed Primers Set, 12 vials of unique dual
indexed primers, 96 reactions

Visit bio-rad.com/CFXDuet for more information about theCFX Duet Real-Time PCR System.
Visit bio-rad.com/PCRplastics and bio-rad.com/PCRreagentsfor more information about PCR and qPCR consumables andreagents, respectively.
BIO-RAD, HARD-SHELL, and MICROSEAL are trademarks of Bio-Rad Laboratories, Inc. incertain jurisdictions. All trademarks used herein are the property of their respective owner.
© 2023 Bio-Rad Laboratories, Inc.
EvaGreen is a trademark of Biotium, Inc.
SYBR is a trademark of Thermo Fisher Scientific Inc. Bio-Rad Laboratories, Inc. islicensed by Thermo Fisher Scientific Inc. to sell reagents containing SYBR Green I for
use in real-time PCR, for research purposes only.

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