page the most prominent real-time PCR cycler are described. In the
cycler descriptions the specifications and the advantages of the
displayed systems are shown. Which real-time platform meets your
requirements best, depends on your research application. Some of the
systems are designed for research with low capacities and others are
for high-throughput applications, most in combination with pipetting
robots. A lot of the displayed cyclers use solid-block for thermal
cycling, other use hot- and cooled-air. Most differences are obviously
the application software, especially in the way of data analysis and
the derived cycle of quantification (Cq is MIQE compliant) crossing
points (CP) or threshold levels (Ct) are computed. Each of
systems employs either one of several general types of fluorescent
for detection. There are also big differences how data are displayed
of the limitations of end-point detection in (RT-) PCR have been
assuaged in real-time PCR systems, a number of which are now on the
market. These systems offer many general technical advantages,
including reduced probabilities of variability and contamination, as
well as online monitoring and the lack of need for post reaction
analyses. Further, some of these systems were
developed with contemporary applications such as quantitative PCR,
HRM (high resolution melting) and high-throughput analysis in mind.
Initial template levels can be
calculated by analysing the shape of the curve or by determining when
rises above some threshold value. A hopefully complete list of the
PCR systems are overviewed on this page and/or summarized below:
Your research. Your
way - QuantStudio™ real-time PCR & digital PCR systems
Flexibility. Versatility. Speed. Precision. Everyone’s needs are unique
and that’s why we have expanded the QuantStudio™ family
of real-time PCR & digital PCR systems. Now you can pick the qPCR
platform that best fits your research requirements - find your fit
today. download brochure
The StepOne™ Real-Time PCR System combines plug and play convenience
and uncompromising performance that is tailor-made for both first-time
users and real-time PCR experts. By utilizing the full range of TaqMan®
gene expression and genotyping assays, the StepOne
System delivers the
kind of high-quality results you've come to expect from the leader in
real-time PCR. The StepOne System provides the power and flexibility
experienced users demand, while its intuitive software guides new users
through experimental design, operation, and analysis. Part of an
elegant, optimized system solution for real-time PCR that includes
software, reagents, and instrumentation, the StepOne System delivers
remarkable performance with remarkable ease-of-use, at a price that can
only be described as...remarkable.
Easy to use software helps you set up your
experiments and get results
Fast runs (<40 minutes) and standard runs (<2
hours) with one
Flexible configurations for PC-free or networked
Preloaded and precalibrated to get you up and
Small, space-saving footprint
Supports real-time PCR applications
* The StepOnePlus™ Real-Time PCR System Upgrade is an
easy upgrade solution for your StepOne™ System. You can go from 3
colors and 48 wells to 4 colors and 96 wells in no time at all.
Biosystems® StepOnePlus™ Real-Time
System, makes it simple and easy to get high-quality real-time PCR
results. This remarkably simple 96 well, 4 color, real-time PCR system
is designed with a user-friendly, yet powerful, interface for
researchers of all experience levels.
Intuitive, flexible software and wizards guide new
their real-time PCR experiments in three easy-to follow steps
4-color/96-well format delivers precise,
quantitative real-time PCR
Long-life LED-based optical system records
FAM™/SYBR® Green, VIC®/JOE™, ROX™ and NED/TAMRA™ dyes for gene
expression analysis, pathogen detection, SNP genotyping, and
One block performs both standard runs in under 2
hours and fast PCR
reactions in less than 40 minutes
Ultra-compact footprint fits any laboratory setting
LCD touchscreen and USB drive provide configuration
enable PC-free operation
Remote monitoring and email notification for
VeriFlex™ Block technology brings six independently
peltier blocks together for precise temperature control and enhanced
StepOnePlus™ system brings advanced real-time PCR technology to a new
level of accessibility. Beginning at the StepOnePlus software homepage,
you can seamlessly navigate through all aspects of the real-time PCR
method including sample and reaction set-up, thermal cycling, and
fluorescent detection. Focused application software analyzes and
interprets experimental results. Depending on the experimental design,
the system can even help you select and order real-time PCR reagents
online by means of convenient links in the Design Wizard.
Remarkable Results - Because
the StepOnePlus system is factory-calibrated for optical and thermal
accuracy, simply remarkable real-time PCR results are available right
out of the box. It can discriminate between 2 populations of 5,000 and
10,000 template copies of a TaqMan® Assay with 99.7% confidence.
and Software Analysis
StepOnePlus System supports any real-time PCR application.
Gene Expression Profiling
Viral Load Analysis
for StepOne and StepOnePlus systems supports a variety of analysis
Relative Standard Curve
Comparative CT (∆∆ CT)
Presence/absence (Plus/Minus) assays with an
internal positive control
Melt curve analysis
Genotyping (including real-time amplification)
software for the Applied Biosystems®
StepOne Real-Time PCR System runs
on the Windows XP® operating system and provides instrument
control, data collection, and data analysis. This user-friendly and
intuitive software package includes the following features:
Experimental design wizards to help you design and
set up experiments.
Pipetting protocols and recipes to quickly set up
Advanced setup for expert users who require
flexibility for more
complex applications, such as multiplexing.
QuickStart setup so that you start a run immediately
and enter plate
information at a later time.
Real-time monitoring of amplification growth curves
enables you to
view run progress.
Remote real-time monitoring so that you can monitor
progress from a
Email notifications to alert you when a run has
started or ended.
Auto-baseline and auto-threshold for simplified data
Automated SNP genotype calling capability with
output and quality-value assignment.
Tool tips for easy identification of sample wells
amplification curves or SNP genotyping plots.
Troubleshooting flags to help you diagnose and solve
Multiple plots view so that you can simultaneously
assess data from
four different perspectives.
Easy cut and paste functionality.
Export easily to PowerPoint, Excel, and graphical
7500 Fast Real-Time PCR System
The Applied Biosystems 7500 FAST Real-Time PCR System is a
versatile, leading edge platform offering enhanced performance
capabilities and a future upgrade to high speed thermal cycling.
Variable excitation and five-color detection enable
you to use a broad range of fluorophores including FAM™/SYBR® Green
I, VIC®/JOE, NED™/ TAMRA™/ Cy3™, ROX™/Texas Red®, and Cy5™ dyes
for precise results
Supports 96-well format plates and 0.2 mL tubes.
Powerful, versatile software including plate set-up
wizards and automated analysis tools make data processing simple and
Advanced data viewing capabilities and automated
analysis tools enable simple and straightforward data processing
The ViiA™ 7 Dx Instrument puts high-productivity qPCR at your
fingertips. With the new OptiFlex™ optical system and 4 interchangeable
block formats from 96-well to 384-well to TaqMan® Array Micro
Fludic Card, the ViiA™ 7 is available for in-vitro diagnostics use.
Proven Applied Biosystems reliability
Using with TaqMan® Array Micro
Fluidic Cards provides an integrated workflow and faster results.
Easy to use—intuitive software,
responsive touch-screen, effortless block exchange without the need for
Ideal for performing medium- to high-throughput real-time
PCR, the ViiA™ 7 Dx Instrument enhances your lab’s productivity.
Quick block changes: Front access
makes it easy to change thermal cycling block formats without having to
move attached peripherals such as robots or computers.
Easy touch-screen interface:
Instrument touch screen provides one-touch protocols for fast and easy
assay setup for a broad range of applications.
Peace of mind: Data recovery
capability for 100+ runs.
The ViiA™ 7 Dx Instrument features the new OptiFlex™
System, which provides enhanced fluorescence detection.
Maximum multiplexing: Six decoupled
excitation and emission filter channels for the greatest number of dye
combinations and maximum multiplexing capabilities.
Flexible data collection: Multiple
ramp method detection formats provide more flexibility for collecting
data during a ramp stage.
Precise quantification: Detect as
small as 1.5–fold changes in target quantities in singleplex reactions.
ViiA™ 7 software drives value in a number of ways,
Intuitive interface and innovative
Convenient walk-away operation
Improved data analysis
Fully compatible with high-throughput
real-time PCR & digital PCR systems
Flexibility. Versatility. Speed. Precision. Everyone’s needs are unique
and that’s why we have expanded the QuantStudio® family of
real-time PCR & digital PCR systems. Now you can pick the qPCR
platform that best fits your research requirements—find your fit today.
The QuantStudio® 6 Flex Real-Time
PCR System is ideal for laboratories with multiple applications
and end users on a limited budget. With a planned upgrade path to a
QuantStudio® 7 Flex System that accommodates automation or
TaqMan® Array Cards, the QuantStudio® 6 Flex System is an ideal
qPCR platform to accommodate changing future needs.
The QuantStudio® 7 Flex Real-Time
PCR System delivers the proven reliability, sensitivity, and
accuracy of the ViiA™ 7 System in a new industrial design. The
QuantStudio® 7 Flex System has been optimized to enable the
broadest range of quantitative PCR applications, with additional dyes,
formats, and automation options.
Combining flexible throughput capabilities with a streamlined workflow,
the QuantStudio® 12K Flex system
takes you from targeted discovery through confirmation and screening,
all on a single platform. Increase your throughput by as much as
32-fold by running 4 OpenArray® plates in a single run on the
QuantStudio® 12K Flex system. It's the equivalent of 32 traditional
384-well qPCR plates or 12,000 data points. When equipped with the
OpenArray® block, a single 9700 thermal cycler and the
QuantStudio® 12K Flex OpenArray® AccuFill™ System, the
QuantStudio® 12K Flex system can produce up to 110,000 data
points or more in an eight-hour day. For those that need the greatest
throughput, add the QuantStudio™ 12K Flex Automation Robot and multiple
9700 thermal cyclers, and you can achieve over 2,000,000 data points in
a day. Start a 12,000 data point experiment typically in 20 minutes or
Now, you can go beyond the limits of real-time PCR with the new QuantStudio® 3D Digital PCR System.
A simple and affordable platform, the QuantStudio® 3D Digital
PCR System produces absolute quantification data with the precision and
sensitivity you need for your more demanding applications. The
QuantStudio® 3D Digital PCR System uses a sealed chip
technology, providing a streamlined, reliable, and robust method for
performing digital PCR. An affordable platform where you can just load
and go—digital PCR is now absolutely attainable.
LightCycler™ from Roche Molecular Biochemicals of
Indianapolis harnesses light technology licensed from Idaho Technology
Inc. of Idaho Falls, and then adds some additional bells and whistles,
such as a built-in microvolume fluorimetric detection system that
enables real-time quantitative PCR. This system employs thin-walled
glass capillary tubes, and typical PCR experiments can be performed in
less than 30 minutes. Currently, this system supports two
fluorescence-based methods for the detection of amplification products:
the "general" DNA stain SYBR Green I (a product of Molecular Probes of
Eugene, Ore.), or sequence-specific hybridization probe pairs. SYBR
Green I exhibits very little fluorescence when free in solution;
emission is greatly enhanced when it binds to the minor groove of the
DNA double helix. Prior to amplification, the reaction mixture contains
the denatured DNA, the primers, and the dye. The low-level background
fluorescence signal generated by the unbound dye molecules is
subtracted during computer analysis. After annealing of the primers, a
few dye molecules can bind to the double strand. During elongation,
more and more dye molecules bind to the newly synthesized DNA,
resulting in dramatically increased light emission. If the reaction is
monitored continuously, this increase in fluorescence can be viewed in
real time. After denaturation of the DNA during the next heating cycle,
the dye molecules are released and the fluorescence signal falls. A
fluorescence measurement is performed at the end of the elongation step
of every PCR cycle to monitor the increasing amount of amplified DNA.
The hybridization probe format employs two specially designed,
sequence-specificoligonucleotides lab-eled with fluorescent dyes. One
oligonucleotide probe carries a fluorescein label at its 3' end;
the other probe carries a different label (LC Red 640 or LC Red 705) at
its 5' end. The chemical nature of the hybridization probes prevents
their extension: one probe contains fluorescein at the 3' end, whereas
the 5'-labeled probe contains a 3' phosphate moiety. The sequences of
the two oligonucleotides are selected so that they hybridize to the
amplified DNA fragment in a head-to-tail arrangement. When the
oligonucleotides hybridize in this orientation, the two fluorescent
dyes are positioned in close proximity to each other. The first dye
(fluorescein) is excited by the LightCycler's light emitting diode
(LED) filtered light source, and emits green fluorescent light at a
slightly longer wavelength. When the two dyes are in close proximity,
the emitted energy excites the dye attached to the second hybridization
probe, which subsequently emits red fluorescent light at an even longer
wavelength. This energy transfer, referred to as fluorescence resonance
energy transfer (FRET), occurs efficiently only when the dyes are in
close proximity (a distance between 1-5 nucleotides). Thus, in this
type of assay, fluorescent intensity measurements are made after the
annealing steps. The increasing amount of emitted fluorescence is
proportional to the increasing amount of DNA generated during the
linear phase of the ongoing PCR process.
Applied Science’s cutting-edge real-time PCR technology
- now available for high-throughput applications
LightCycler 480 Real-Time PCR System is a fully integrated
multiwell-plate based real-time PCR platform for highly accurate
qualitative and quantitative detection of nucleic acids. Building on
the benefits of Roche's capillary-based LightCycler® Systems, it
goes one step further in offering enhanced throughput, compatibility
with automation equipment and maximum flexibility regarding hard- and
Providing novel ways to combine speed and accuracy without compromises,
the LightCycler® 480 Real-Time PCR System meets the needs of a
broad range of applications in research fields such as gene expression
studies, discovery and analysis of genetic variation or array data
LightCycler® 480 Real-Time PCR System benefits
Benefit from novel thermal cycling and data
capture technologies to achieve outstanding temperature homogeneity and
Increase the sensitivity and yield of your
real-time PCR applications, with robust LightCycler® 480 reagents
and specially engineered clear or white consumables.
Analyze true raw data, breaking free from the need
to use passive reference dyes or normalization plates to get accurate
Flexibly switch between 96- or 384-well plate
formats, exchanging thermal block cyclers in just a few minutes with no
Select from numerous assay formats and detection
dyes, expanding the range of real-time PCR applications in your lab.
Benefit from the intuitive and user-friendly Roche
Take advantage of the automation capabilities of
the LightCycler® 480 System, integrating it as part of a LIMS in
combination with robotic pipetting and plate loading.
Conveniently analyze gene expression or genotyping
data using a broad range of basic and advanced methods.
Know that your data are protected, using a system
compatible with 21 CFR part 11 requirements
Gene Scanning by High Resolution
Analysis generally requires the use of:
a special generic DNA dye
that works at high, saturating concentrations without inhibiting PCR
and therefore leads to homogeneous staining of homo-or heteroduplex DNA
instrument with suitable excitation/emmission wavelengths, high data
acquisition rates, and outstanding temperature homogeneity
a software algorithm that analyzes the shape of the
melting curves and groups those that are similar.
In a Gene Scanning experiment, sample
first amplified via
real-time PCR in the presence of a proprietary saturating DNA dye. A
melting curve is then performed using high data acquisition
rates, and data are finally analyzed using a Gene Scanning Software, by
three basic steps:
Normalization:the pre-melt (initial
fluorescence) and post-melt
signals of all samples are set to uniform, relative values from 100% to
temperature axis of the normalized melting curves is shifted to the
point where the entire double-stranded DNA is completely denatured.
Samples with heterozygous SNPs can then be easily be distinguished from
the wild type by the different shapes of their melting curves.
the differences in melting curve shape are further analyzed by
subtracting the curves from a reference curve. This helps cluster
samples automatically into groups that have similar melting curves
(e.g., those who are heterozygote as opposed to homozygotes).
High-resolution melting curve analysis on
the LightCycler 480 PCR system (presented by
Roche Aplied Science)
Science´s LightCycler® family of real-time PCR systems offer
fast, accurate and versatile platforms for genetic
variation research. The new plate-based LightCycler® 480 System
provides the temperature
homogeneity and optical characteristics required for high-performance
melting-curve analysis (MCA). On
the level of data acquisition and available detection channels, this
new instrument opens the way to more
advanced applications in the emerging field of gene scanning where
amplicons can be screened for
unknown sequence variations with low efforts in time and cost. The LightCycler® 480
real-time PCR system: a versatile platform for genetic variation
Real-time PCR is a well established technique for studying genetic
variation using various probe-based methods for genotyping as well as
high-resolution analysis of whole amplicons melted in the presence of
saturating DNA dyes. The latter, relatively new, method allows
screening for unknown mutations or DNA modifications. The
LightCycler® 480 real-time PCR system is a multiwell plate–based
instrument that provides integrated applications for detecting and
characterizing genetic variation using all these methodological
to HRM-assays on the LightCycler 480 System- Examples
High-resolution melting curve analysis (hrMCA) is an attractive
technique to scan for unknown mutations in genes. To evaluate how easy
or difficult it is to design hrMCA assays using the LightCycler®
480 Instrument, we selected 3 different fragments in exon 11 of the
BRCA1 gene, designed an MCA assay, and tested its sensitivity to detect
Large-scale, PCR-based gene analysis can only be performed if signals
are generated, captured and analyzed in a highly reproducible manner.
More precisely, thermal control, optical readout and the algorithms
used to characterize genes qualitatively and quantitatively have to be
highly precise and reliable. The LightCycler® 1536 System from
Roche Applied Science meets these requirements. The novel platform,
which utilizes a proprietary 1536-well plate, is capable of performing
high-speed, qPCR-based DNA/RNA analyses in an array-like format.
Throughput — Redefined
Break through to a new era in high-throughput real-time PCR with the
powerful LightCycler® 1536 System, precisely engineered for
miniaturization and parallelization.
Increase throughput fourfold - Generate 1,536 data
points in a single run in less than 50 minutes.
Make the most of your resources - Use significantly
less reagent and sample with reaction volumes of just 0.5-2.0 µl.
Generate PCR data you can trust - Ensure research
continuity with this extension of proven plate-based technology.
Easily integrate into your existing workflow - Take
advantage of this streamlined system, specifically designed for
automation and high-throughput.
For life science research only. Not for use in diagnostic
A small yet complete system with
all features you've come to expect from LightCycler® Instruments
1. The LightCycler® Nano Instrument produces high
target specificity with high sensitivity characteristic of all
2. Fast protocols use state-of-the-art, 32-well thermal
cycler technology for excellent thermal precision.
3. Robust solid-state optics provide complete spectral
Sleek, built to enjoy in every
lab, on every bench
1. Small enough for plenty of space on your benchtop.
2. Experience ease-of-use with flexible workflows.
3. Easily install using plug and play.
4. So quiet. You won't know it's running if it wasn't for
the light show.
5. Intuitive software from assay setup to analysis.
Great looks meets fast,
sensitive and accurate qPCR
1. The LightCycler® Nano System embodies experimental
2. The instrument uses standard 8-tube PCR strips that
come with caps.
3. Flexible reaction volumes permit different levels of
4. Proven reagents for SYBR Green or probe-based assays
5. Use different methods for quantification and mutation
analysis, including HRM.
6. Use your preferred operating system, Mac OS or Windows,
or go standalone with a USB flash drive.
LightCycler® 96 Instrument is a real-time PCR system for
rapid cycling up to 96 samples. Applications include absolute and
relative quantification, melting curve analysis, and endpoint
genotyping. Precise LightCycler® 96 Instrument thermal homogeneity
cycling speed produce accurate and reproducible results in a very short
The LightCycler® 96 Instrument’s optical detection system flexibly
detects sequence-dependent probes, such as hydrolysis probes and
sequence-independent dyes (i.e., SYBR Green I). Multiplex and
multicolor capabilities permit the use of up to four different
fluorescent dyes. New innovative LightCycler® 96 Application and
Instrument Software creates a simple analysis workflow with easy import
and export functions, email notifications after each run, online
monitoring flexibility, and server-based network interfacing.
Plate-based real-time PCR system with 96-well
Temperature gradient (assay optimization)
Standalone system (integrated PC)
Touchscreen with separate instrument software
(free to load on any PC)
PC analysis software can be loaded free of
charge on any PC
Obtain fast, highly sensitive,
and reproducible results for qualitative and quantitative detection of
nucleic acids and SNP analysis with the versatile, plate-based
LightCycler® 96 System:
Benefit from a cutting-edge, low-mass thermal
that offers excellent well-to-well homogeneity for a broad range of
temperatures, and access to gradient features. The electroforming
production process allows a perfect fit of the 96-well plate (negative
structure) to the block (positive structure, formed by a galvanic
reaction) and avoids air cushions in between both.
Maximize sensitivity and speed using novel
(combining sturdy glass fibers with high-intensity broad-spectrum
LEDs), and obtain simultaneous detection of all wells within
Install the software on any PC for free, with
no license requirements, and run the system according to your specific
Standalone, using a flash drive to transfer
Connected to a PC
Connected to a PC network via Ethernet
Experience advanced software features, such as
real-time remote monitoring, e-mail notification, heat-maps, wizards,
and many more options to operate the intuitive and easy-to-use software.
Extract meaningful data via state-of-the-art
analysis methods, then easily export results in a ready-to-publish
Choose the optimal type of LightCycler®
Plate or 8-Tube strips (white, recommended) or clear to best fit your
assay format, application, and workflow.
This instrument was the world’s first rapid cycling PCR machine and was
a collaboration with the University of Utah and Idaho Technology. While
not a real-time PCR instrument itself, the success of its rapid cycling
technology led to the original LightCycler® with thousands of
air-based rapid qPCR machines still in use today. For more info on the
theory behind rapid cycling see this link http://dna.utah.edu/RapidPCR/Top_Rapid_PCR.html
The RapidCycler 2 uses 1.5 mm Roche LightCycler® capillary tubes
which are ideally suited for high-speed temperature cycling due to
their thin walls and extremely high surface-are-to-volume ratio.
By combining glass capillary tubes and air as the cycling medium we
have cut sample temperature lag to less than a second, therefore
yielding high temperature ramp rates. Inside RapidCycler's cylindrical
reaction chamber, tornado-like conditions ensure both temperature
uniformity and rapid heat exchange within the sample. this allows the
RapidCycler to spend less than one second at the high and low
temperatures and still be certain that each sample has reached the
The original LightCycler was licensed to
Roche and is no longer sold by
For use with Idaho Technology's LightCycler Instrument only
Contact us for support or information.
Instructions: To obtain a copy of our latest LightCycler 24 & 32
software, contact us directly by e-mail or phone (801) 736-6354 x 349.
Please have the serial number of your machine ready.
For a history of the exciting University of Utah / Idaho Technology
partnership that brought you rapid real-time PCR in the same year as
the “big guys” see this link. http://dna.utah.edu/LightCycler/Top_LightCycler.html
realplex - real-time PCR solutions
has developed a real-time PCR system that meets the requirements of the
latest quantitative PCR applications. Comprehensive yet intuitive
software paired with recent advances in real-time PCR technology make
the Mastercycler ep realplex a critical
to modern science and research. The Mastercycler ep realplex features
the latest technology, united with good design and a quality finish.
A minimal number of moving parts in the optical detection module
the vulnerability of the overall system and promotes consistently high
sensitivity in your realtime PCR experiments. The overall system has
been solidly engineered, to ensure quiet operation in the laboratory.
Real-time without limits
from two thermomodules depending on your requirements. The "standard"
variant is fitted with a specially finished 96-well aluminum block,
while the "high-speed" variant has a 96-well silver block with Impulse
PCR and extremely fast heating and cooling rates. Regardless of the
thermomodule selected, reproducible results can be attributed to the
highest accuracy and uniformity of Mastercycler ep thermal cycling.
Both thermomodules can be combined with two different optical modules, realplex2
and realplex4, which permit the use of nearly all
fluorescent dyes used in real-time PCR. In order to give you the
possible flexibility in experimental design, Eppendorf's realplex
a completely open system concept. Pre-established assays optimized for
real-time PCR reagents and your preferred tubes or plates can be easily
transferred to the Mastercycler ep realplex. Technical
fluorescent dyes chosen for each experiment are excited by an array of
96 individual LEDs, which have a substantially longer lifespan in
contrast to halogen lamps. The result is a real-time PCR system that
is less susceptible, minimizing equipment downtime and the frequency
of maintenance and associated delays to your experiments.
The LEDs generate a blue
light at a wavelength of ~470 nm, which can excite nearly all
fluorophores used in realtime PCR (including SYBR®Green, FAM, VIC,
TET, HEX, ROX, JOE, TAMRA). Emitted fluorescence is focused through an
array of lenses and is passed to the optical detection unit through 96
individual optical fibers.
Immediately upstream of the
detectors, two high-quality emission filters with bandwidths of 10 nm
nm are contained in a filter rotor.
New type channel
photo-multipliers (CPM) serve as the detectors, which in contrast to
conventional photomultiplier tubes, are far less sensitive to magnetic
Mastercycler ep realplex
Smart Workstations epMotion® 5070 and 5075
operation with a
operator convenience due to a comprehensive database of validated
applications and consumables
of 1 µl - 1,000µl volumes
optical sensor detects tubes and plates,as well as their liquid levels
and pipette tips