PRODUCT BROCHURE Real-Time PCR Systems

StepOne and StepOnePlus ™ ™

Real-Time PCR Systems

Remarkably Simple Systems. Simply Remarkable Results.
Step Up To High Performance Real-Time PCR

Remarkably Simple Systems

Applied Biosystems, a leader in technology solutions for life science research, introduces
its latest innovations in real-time PCR—the StepOne™ (48-well) and StepOnePlus™ (96-well)
Real-Time PCR Systems. These remarkably simple real-time PCR systems are specifically
designed with a user-friendly, yet powerful interface for both new and experienced
real-time PCR researchers.
 Features At A Glance

StepOne System StepOnePlus System

Throughput/Wells 48 96
™ ®
FAM /SYBR Green dyes ✓ ✓
®
VIC /JOE dyes™
✓ ✓

ROX dye ™
✓ ✓

NED/TAMRA dyes ™
✓

Figure 1. Software Homepage VeriFlex™ Block ✓

System Features
The StepOne™ and StepOnePlus™ Systems bring advanced
real-time PCR technology to a new level of accessibility—even
System Highlights
for first-time users. Both systems measure amplification as
For both novice and experienced researchers, the StepOne or
it occurs, cycle by cycle, allowing for precise and quantitative
StepOnePlus Systems have what you need.
measurements during the exponential phase of PCR. Beginning
at the homepage (Figure 1), you can navigate seamlessly • Cost-effective, 3-color/48-well (StepOne) or 4-color/96-well
through all aspects of the real-time PCR process including (StepOnePlus) Systems deliver precise, quantitative real-time
sample and reaction set-up, thermal cycling, and fluorescent PCR results
detection. Focused application software analyzes and interprets • Long-life LED-based optical system records fluorescence
experimental results. Depending on the experimental design, ®
from FAM™/SYBR® Green I, VIC /JOE™, and ROX™ dyes for
the system can even help you select and order real-time PCR gene expression analysis, pathogen quantitation, SNP
reagents online by means of convenient links in the software genotyping, and presence/absence assays; Note: The
(optional). The system provides two-fold discrimination with StepOnePlus System also accommodates the TAMRA™ dye
99.7% confidence, along with 9 logs of dynamic range. Highly
sensitive, these systems detect 10 copies of RNaseP in a 30-μL
• Both systems perform standard and fast PCR reactions in
reaction. Additionally, for laboratories with the StepOne System less than 40 minutes
that require greater throughput, an upgrade kit is available.* • Ultra-compact footprint fits any laboratory setting
StepOne Upgrade • LCD touchscreen and USB drive provide configuration
For those customers whose lab needs are changing, Applied flexibility and enable PC free operation
Biosystems offers the StepOne System upgrade. Just send
• Remote monitoring and email notification for convenience
in your existing system and you’ll receive a StepOnePlus
and time-savings
instrument until your upgraded system arrives in your lab.
• The StepOnePlus System features VeriFlex™ Block
Easy-to-Use Experimental Design Wizard
technology, which combines six independently
Both systems provide an experimental Design Wizard to
controllable peltier blocks for enhanced PCR
assist you in setting up your first real-time PCR experiment.
functionality and precise temperature control
Just click on the Design Wizard and start answering questions
about the real-time PCR experiment you wish to perform. The • The StepOne System is upgradeable to the StepOnePlus
Wizard walks you through quantitation methods, detection System to meet your changing research needs
chemistry, ramp speeds, and nucleic acid template types.
From standards to plate layout, the Design Wizard guides the
entire process, including reaction mix and standard dilution
calculations. You’re now ready to explore the simply remarkable
features of these systems.

*StepOnePlus System Upgrade Kit coming soon

Simply Remarkable Interface
Flexibility The StepOne and StepOnePlus Systems can be installed in
Both the StepOne™ and StepOne Plus™ Systems adapt to multiple distinct configurations, providing unmatched flexibility
almost any workflow with flexible instrument control and data and convenience. The unique standalone (PC free) configuration
management. From the touchscreen control panel, you can provides an ultra-compact footprint that will fit into any labora-
quick-start your experimental run without PC connectivity. You tory. A direct connection to a LAN enables remote monitoring
can also create a new protocol, view the history of your last of experimental progress and downloading of the completed
run, or see protocol details. When the run is complete, data run file to the PC at your desk.
can be downloaded easily onto a USB flash stick or saved
System Configurations (Figure 4)
to a PC. Either system can be connected directly to a Local
1. PC controlled
Area Network (LAN), and you can monitor the progress of the
experiment, send new instructions to the system, download 2. PC free
data, and edit the instrument profile. The software for the two 3. Networked
systems also contains a convenient email feature that notifies
you when your experiment is complete and ready for analysis. 4. PC controlled connected to LAN

5. PC controlled with networked instrument

Figure 2. Browse Experiment Page On Touchscreen Figure 3. Run Monitor Screen On Touchscreen

Figure 4. Five Distinct Configurations

1. PC controlled 2. PC free 3. Networked

LAN
Software • Tools for easy identification of sample wells when
The StepOne™ and StepOnePlus™ Systems software contains viewing amplification curves or SNP genotyping plots
unique features not available in other real-time PCR instru-
ments. For example, data can be analyzed from multiple
• Email notifications to alert you when a run has started
perspectives in the Multiple Plots view (See Figure 5). The or ended
software constructs four-plot, side-by-side views of all data • Easy cut-and-paste functionality
aspects including the amplification plot, standard curve,
• Export easily to PowerPoint®, Excel®, or directly as a .jpeg file
multi-component data plots, and raw data. It also displays this
data next to the plate layout for easier analysis. Another novel
Figure 5. StepOne System Multiple Plots View
feature is the software’s ability to automatically identify wells
that might compromise the success of an experiment. During
data analysis, the software generates a quality-control report
table that flags wells based on criteria such as amplification
in a negative-control well, the absence of a signal in a well, or
a high CT standard deviation in a replicate group. This feature
reduces analysis time and allows even researchers new to
real-time PCR to have confidence in their results. Furthermore,
you can customize this feature by disabling or modifying
quality flag settings to suit your experimental needs.

Software Highlights
• Experimental Design Wizards to help you design and set
up experiments
• Pipetting protocols and recipes to set up
experiments quickly
• Advanced set-up for expert users who require flexibility Figure 6. StepOne System Reaction Set-up View
for more complex applications, such as multiplexing
• Quick-start set-up so you can begin a run immediately
and enter plate information later
• Real-time monitoring of amplification growth curves enables
you to view run progress (can be viewed from a remote PC)
• Auto-baseline and auto-threshold for simplified data analysis
• Multiple Plots view for simultaneous data assessment from
four perspectives
• Automated SNP genotype-calling with intuitive graphical
output and quality-value assignment
• Troubleshooting flags to help you diagnose and solve
problematic experiments

For each target in the reaction plate, the StepOne System software lets you
review and edit calculated volumes for preparing standards and PCR reactions.

4. PC controlled connected to LAN 5. PC controlled with network instrument

USB—10/100 Adapter
LAN
Simply Remarkable Applications
Instrument Chemistries • Melt curve analysis
Both StepOne Systems are equipped with fluorophore
• Genotyping (including real-time amplification)
detection chemistries that include FAM™ and VIC® dye-labeled
TaqMan® MGB probe-based assays, VIC and TAMRA™ dye-
Applications
labeled probe-based assays and SYBR® Green I dye chemistry.
The StepOne Systems support any real-time PCR
[Note: the TAMRA dye is available only with the StepOnePlus
application. Predesigned or custom assays exist for the
System.] TaqMan probe-based assays provide outstanding
following applications:
specificity and sensitivity, and SYBR Green I dye chemistry is an
economical alternative for target identification, initial screening • SNP Genotyping • Translocation Analysis
assays, or assays that require only a few reactions. • Gene Expression Profiling • Gene Detection
Software Analysis • MicroRNA Expression • Viral Load Analysis
The software for the StepOne Systems supports a variety
of analysis methods, including: For information about existing Gene Expression, MicroRNA and
Translocation Analysis Assays, please visit www.allgenes.com
• Absolute Quantitation
- Standard curve SNP Genotyping Assay information can be found at
www.allsnps.com
• Relative Quantitation
- Relative standard curve Speed
- Comparative CT (ΔΔCT) The StepOne and StepOnePlus Systems perform both
standard and fast thermal cycling on the same block with no
• Presence/absence (plus/minus) assays with an internal modification. Standard thermal cycling requires less than two
positive control

Figure 7. Broad Dynamic Range Figure 8. Genotyping Analysis

Amplification Plot Allele Discrimination Plot (SNP Assay: SNP Assay 1)

10

4.1

3.6

3.1

2.6
1
Allele 2
ΔRn

2.1

1.6

1.1

0.6
0.1

0.1

0.0 0.2 0.4 0.6 0.8 1.0 1.2 1.4 1.6 1.8 2.0
2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40 Allele 1
Cycle
Homozygous 1/1 Heterozygous 1/2 Homozygous 2/2 Undeterminded

Amplification plot shows log of change in normalized reporter fluorescence StepOne and StepOnePlus software automatically determines genotypes and
plotted vs. PCR cycle number. This plot from the StepOne™ System illustrates 9 generates an intuitive graphic representation of results in a cluster plot report
logs of linear dynamic range for a TaqMan® Assay of cDNA containing the 18S that helps you view data across populations or samples. Results are from human
target sequence in tenfold serial dilutions. CYP2C19*2 TaqMan ® SNP Genotyping Assay (using the StepOnePlus System).
hours, and fast thermal cycling significantly reduces the run
times of quantitative real-time PCR applications by delivering
results in 40 minutes. Fast cycling is ideal for maximizing the
number of runs on an instrument in any given work day.

VeriFlex™ Blocks
The 96-well StepOnePlus System fea-
tures VeriFlex™ Block technology, which
brings six independently controllable
peltier blocks together for precise
temperature control and enhanced
PCR functionality. VeriFlex Blocks
deliver flexibility for those who have
probes and primers that are optimized
at different annealing temperatures. VeriFlex Block (96-well)

Results
Because the StepOne and StepOnePlus Systems are factory-
calibrated for optical and thermal accuracy, simply remarkable
real-time PCR results are available right out of the box. Both
systems can discriminate between two populations of 5,000
and 10,000 template copies of a TaqMan® assay with 99.7%
confidence. Both systems also demonstrate a linear dynamic
range of 9 log units or more, as shown by the amplification
plot (See Figure 7).

Figure 9. Melt Curve Analysis Figure 10. Multiplex Assay

Melt Curve Multiplex Assays

10

3.4

2.9

2.4
1
Derivative Reporter (-Rn’)

1.9
ΔRn

1.4

0.1
.9

.4

- .1

0.01

65.0 70.0 75.0 80.0 85.0 90.0 95.0 0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40 42

Tm: 79.2 Cycle
Temperature (°C)

In this experiment, standard PCR was run to generate amplicons for 18S, Multiplex TaqMan ® Assays on the StepOnePlus™ System showing
ß-actin and GAPDH. Aliquots of each amplicon were combined into one amplification of cDNA (96 samples) using probes labeled with VIC ®
reaction tube and a melt curve analysis was performed. and FAM ™ reporters for 18S and TGF-ß target sequences respectively.
Systems At A Glance

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StepOne™ System StepOnePlus™ System

• 48 wells for lower throughput • 96 wells for higher throughput

• 3 color for basic applications research • 4 color for more flexibility

• Easily upgradeable to StepOnePlus System • VeriFlex™ Block technology for thermal cycling flexibility
The Remarkable
Applied Biosystems Solution

Applied Biosystems offers a complete range of real-time types including animal and plant tissue, cultured cells, blood,
PCR reagents and design/analysis software to assist bacteria, and yeast. Blood, bacteria, and formalin-fixed material
both experienced and novice users of the StepOne™ and require RNA isolation kits designed specifically for that sample
StepOnePlus™ Real-Time PCR Systems. During experimental type, whereas most eukaryotic samples can be processed with
set-up, the Design Wizard automatically creates a material excellent results using Ambion’s standard RNA isolation kits.
list that links directly to the Applied Biosystems Store for
TaqMan® Gene Expression Master Mix
convenient ordering of real-time PCR master mixes and
Tailored for quantitative real-time PCR experiments, the
consumable plasticware (optional).
TaqMan Gene Expression Master Mix delivers robust
Service and Support performance for both routine and challenging quantitative
Purchase of either of these systems includes a limited warranty applications. It enables specific target detection across a large
on parts and labor.* Depot repair and update services are dynamic range with high sensitivity down to a single copy
available for the StepOne and StepOnePlus Systems through number. Furthermore, it offers duplex capability and reproduci-
Applied Biosystems. Technical support and service are provided bility at less than two-fold discrimination.
worldwide. Additionally, an upgrade is available from the
TaqMan® Genotyping Master Mix
StepOne to StepOnePlus System.
Specifically formulated for reliable, cost-effective detection of
Applied Biosystems Training Programs single nucleotide polymorphism (SNP) detection, the TaqMan
Theoretical and practical hands-on laboratory sessions provide Genotyping Master Mix provides accurate and reproducible
in-depth instruction for customers seeking to gain proficiency allelic discrimination with well-separated clusters for
and streamline workflows with Applied Biosystems solutions. exceptional call rates, premium performance in challenging
Applied Biosystems Application Support Centers offer genotyping assays, and excellent benchtop stability for
interactive, computer-based training, fully-staffed working superior flexibility to meet various throughput needs.
laboratories, and the unique opportunity to collaborate with
TaqMan® MicroRNA Assays
Applied Biosystems scientists and product development teams.
Applied Biosystems offers TaqMan MicroRNA Assays to
Many hands-on courses can also be conducted at the customer
quantify microRNAs with the sensitivity and specificity of
site. Regional training and live and self-paced web-based
TaqMan Assay chemistry. MicroRNA Assays are available for
courses are also available.
human, mouse, rat, Arabidopsis, Drosophila, and C. elegans.
TaqMan® SNP Genotyping Assays
Primer Express® Software
The TaqMan® SNP Genotyping Assays collection includes
Applied Biosystems Primer Express Software v3.0 facilitates
over 4.5 million pre-designed human and mouse genome-wide
primer design with TaqMan probes for real-time PCR or
assays of which 2.5 million are HapMap SNPs, 70,000 cSNPs,
endpoint PCR analyses. It is also ideal for primer design using
160,000 validated assays, and over 2,500 Drug Metabolism
SYBR® Green I dye chemistries. The Primer Express Software
Genotyping Assays. Additionally, our Custom TaqMan® SNP
ships with the StepOne and StepOnePlus Systems.
Genotyping Assays service lets you create your own custom
assays by submitting target SNP sequences for any genome. For more information about the products, reagents,
consumables, assays and kits listed on this page, please
Ambion and Applied Biosystems RNA Isolation Kits
visit info.appliedbiosystems.com/steponeplus
Ambion and Applied Biosystems offer a range of RNA
isolation kits that are suitable for a wide variety of sample

*Warranty length varies by region

TaqMan Assays ®

Applied Biosystems offers the most comprehensive set of inventoried TaqMan® Gene Expression and
SNP Genotyping Assays available. More than 700,000 Gene Expression Assays and over 4.5 million
pre-designed human, and 10,000 pre-designed mouse SNP Genotyping Assays are available at your
fingertips. Alternatively, you can submit your target DNA sequence from any organism, and we’ll custom-
build an assay for you. Applied Biosystems also offers TaqMan® MicroRNA Assays to quantify miRNA
with the sensitivity and specificity of TaqMan® Assay chemistry. For more information on gene expression
assays, visit www.allgenes.com; for information on SNP Genotyping Assays, visit www.allsnps.com

TaqMan® Gene Expression, SNP Genotyping, and MicroRNA Assays

TaqMan® Assays Selection Guide Application

Gene Expression* SNP Genotyping† MicroRNA‡

TaqMan® Pre-designed Assays Yes Yes Yes

(Inventoried and Made-to-Order)

Custom TaqMan® Assays Yes Yes No

Species Number of Inventoried and Made-to-Order Assays

Human > 2,024,000 > 4,000,000 > 300

Mouse > 179,000 > 10,000 > 240

Rat > 128,000 § > 180

Drosophila melanogaster > 38,000 § > 50

Arabadopsis thaliana > 95,000 § > 40

Caenorhabditis elegans > 90,000 § > 60

Canine > 6,000 § N/A

Rhesus macaque > 70 § N/A

* Includes miRNA, gene copy number, and mitochondrial assays

† Includes HapMap and drug metabolism genotyping assays
‡ Gene expression only
§ Custom TaqMan Assays are available for any SNP, transcript, and genome.

Figure 11. The reagents ordering page in the experimental

Design Wizard
Reagents and Disposables

®
A complete line of reagents, including TaqMan® Fast Universal PCR Master Mix, TaqMan Universal PCR
®
Master Mix, Power SYBR Green PCR Master Mix, and disposables, including 96-well plates, is available
for use with the StepOne™ and StepOnePlus™ Real-Time PCR Systems. These products can easily be
added to a shopping list for future reference or for ordering through the “Materials List” link in the
experimental Design Wizard (See Figure 11).

Select Reagents, Consumables, and Service Contract Offerings for Your

Applied Biosystems StepOne and StepOnePlus Real-Time PCR System

Category/Product Description Quantity P/N

Seals and Covers

MicroAmp™ 48-Well Optical Adhesive Film 25 films 4375928

MicroAmp™ 48-Well Optical Adhesive Film 100 films 4375323

MicroAmp™ 96-Well Optical Adhesive Film 25 films 4360954

MicroAmp™ 96-Well Optical Adhesive Film 100 films 4311971

Reaction Plates

MicroAmp™ Fast Optical 48-Well Reaction Plate 20 plates 4375816

MicroAmp™ Fast Optical 96-Well Reaction Plate with Barcode (0.1 mL) 20 plates 4346906

MicroAmp™ Fast Optical 96-Well Reaction Plate with Barcode (0.1 mL) 200 plates 4366932

8-Well Strips

MicroAmp™ Fast 8-Tube Strip, 0.1 mL 125 strips 4358293

MicroAmp™ Optical 8-Cap Strip 300 strips N8010535

Accessories

MicroAmp™ Fast 48-Well Tray 10 trays 4375282

MicroAmp™ 96-Well Tray/Retainer Set for Veriflex™ Block Systems 10 trays 4381850

MicroAmp™ 48-Well Base Adaptor 5 adaptors 4375284

Reagents

Power SYBR® Green PCR Master Mix 5mL 4367659

TaqMan® Gene Expression Master Mix 5mL 4369016

TaqMan® Genotyping Master Mix 10mL 4371355

TaqMan® Fast Universal PCR Master Mix 2 x 1.25 mL 4352042

TaqMan® One-Step RT-PCR Master Mix Reagents Kit 5mL 4309169

High Capacity cDNA Reverse Transcription Kit 200 reactions 4368814

Service Plan

For more information contact your local sales representative SCSTEPONE

ORDERING INFORMATION

Description Part Number

StepOne™ Real-Time PCR System 4376357

™
StepOne Real-Time PCR System with Laptop Computer 4376373

StepOne™ Real-Time PCR System with Tower Computer 4376374

StepOnePlus™ Real-Time PCR System 4376600

StepOnePlus™ Real-Time PCR System with Laptop Computer 4376598

StepOnePlus™ Real-Time PCR System with Tower Computer 4376599

™
StepOnePlus Real-Time PCR System Upgrade Kit* 4379216

*For users of the StepOne™ Real-Time PCR System. Coming soon.

To learn more about the new StepOne and StepOnePlus Real-Time PCR Systems
from Applied Biosystems, contact your local sales representative, or visit us on the
Web at info.appliedbiosystems.com/steponeplus

For Research Use Only. Not for use in diagnostic procedures.

NOTICE TO PURCHASER:
The StepOne™ and StepOnePlus™ Real-Time PCR Systems are covered by one or more of US Patents Nos. 5,038,852, 5,333,675, 5,656,493, 5,475,610, 5,602,756, 6,703,236,
6,814,934, and corresponding claims in their non-US counterparts, owned by Applera Corporation. No right is conveyed expressly, by implication, or by estoppel under any other
patent claim, such as claims to apparatus, reagents, kits, or methods such as 5’ nuclease methods. Further information on purchasing licenses may be obtained by contacting the
Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA.
© 2007 Applied Biosystems. All rights reserved. Applera, Applied Biosystems, AB (Design), and VIC are registered trademarks and FAM, JOE, NED, ROX, StepOne, StepOnePlus,
TAMRA and VeriFlex are trademarks of Applera Corporation or its subsidiaries in the US and/or certain other countries. Ambion is a registered trademark of Ambion, Inc., an
Applied Biosystems Business.
TaqMan is a registered trademark of Roche Molecular Systems, Inc.
SYBR is a registered trademark of Molecular Probes, Inc.
All other trademarks are the sole property of their respective owners.
Printed in the USA, 06/2007 Publication 117BR08-02

Headquarters International Sales

850 Lincoln Centre Drive | Foster City, CA 94404 USA For our office locations please call the division
Phone 650.638.5800 | Toll Free 800.345.5224 headquarters or refer to our Web site at
www.appliedbiosystems.com www.appliedbiosystems.com/about/offices.cfm