Download Custom RT-qPCR Assays and Panels for any Human, Mouse, or Rat

RealTime ready
Custom RT-qPCR Assays and Panels
for any Human, Mouse, or Rat Gene
RealTime ready qPCR Assays and Panels
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RealTime Ready Configurator
Search by Focus Panel
Search by Pathway
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Ordering Information
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For life science research only.
Not for use in diagnostic procedures.
License Disclaimer:
A license to perform the patented 5’ Nuclease Process for research is obtained by the
purchase of (i) both Licensed Probe and Authorized 5’ Nuclease Core Kit, (ii) a Licensed
5’ Nuclease Kit, or (iii) license rights from Applied Biosystems.
This product contains Licensed Probe. Its purchase price includes a limited, non-transferable immunity from suit under U.S. Patent No. and 5,804,375 (claims 1-12 only (exp.
9/8/2015)) and corresponding patent claims outside the United States. The purchase of
this product includes a limited, non-transferable immunity from suit under the foregoing
patent claims for using only this amount of product for the purchaser’s own internal research. The right to use this product in the 5’ Nuclease Process under the corresponding
patent claims outside the United States, can be obtained through purchase of an Authorized 5’ Nuclease Core Kit. Except under separate license rights available from Applied Biosystems, no right under any other patent claim, or to perform commercial services of any
kind, including without limitation reporting the results of purchaser’s activities for a fee or
other commercial consideration, or to sublicense, repackage with other products, or resell in any form, is conveyed expressly, by implication, or by estoppel. This product is for
research use only. Diagnostic uses under Roche patents require a separate license from
Roche. Further information on purchasing licenses may be obtained from the Director of
Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA.
ProbeLibrary is covered by US and other patent applications owned by Exiqon A/S.
Locked Nucleic Acids (LNA) are covered by U.S. Patents No US 6,794,499 (exp. 9/11/2018),
US 6,670,461 (exp. 9/11/2018), US 6,268,490 (exp. 3/9/2018) & US 6,770,748 (exp. 3/9/2018)
and other patents and patent applications owned by Exiqon A/S and Prof. Takeshi Imanishi.
The quenche Parts of the Software used for the LightCycler® 480 System are licensed
from Idaho Technology Inc., Salt Lake City, UT, USA. This product is covered by one or
more of U.S. 6,197,520 (exp. 8/13/2019), 6,303,305 (exp. 3/30/2019), 6,387,621 (exp. 9/8/2019),
6,503,720 (exp. 2/20/2021), 6,730,501 (exp. 3/13/2022) and corresponding claims in their
non-U.S. counterparts, owned by Roche Diagnostics GmbH and/or licensed from Idaho
Technology, Inc.
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This LightCycler® 480 Real-Time PCR System is a real-time thermal cycler licensed for
research use only under U.S. Patent No. 6,814,934 (exp. 5/2/2011) and corresponding
claims in its non-U.S. counterparts, and under one or more of U.S. Patents Nos. 5,475,610
(exp. 12/12/2013), 5,602,756 (exp. 3/14/2011), 6,703,236 (exp. 11/29/2010), 7,238,517 (exp.
11/29/2010), 7,504,241 (exp. 11/29/2010), 7,537,377 (exp. 2/9/2020) or corresponding
claims in their non-U.S. counterparts, owned by Applied Biosystems, LLC. No rights are
conveyed expressly, by implication or estoppels to any other patent claims. The LightCycler® 480 Real-Time PCR System is not intended for in vitro diagnostic use in the U.S. For
further information on purchasing additional rights, contact the Director of Licensing at
Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California, 94404, USA.
No right is conveyed expressly, by implication, or by estoppels under any other patent
claim, such as claims to apparatus, reagents, kits, or methods such as 5’ nuclease methods. License rights to practice PCR methods under the foregoing Roche patents for research and other non-in vitro diagnostic applications may be purchased from Applied Biosystems or may be obtained by purchasing licensed reagents from Roche, Applied
Biosystems, or other Authorized third party. For information on purchasing licenses for research and other non-in vitro diagnostic applications, contact the Director of Licensing at
Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA.
A license to practice PCR methods with real-time detection under patents of F. HoffmannLa Roche Ltd, and Roche Molecular Systems (“Roche”) for in vitro diagnostic applications
may be purchased from Roche or may be obtained by purchasing in vitro diagnostic reagents from Roche or any other Authorized third party. The LightCycler® 480 Real-Time
PCR System is not intended for in vitro diagnostic use in the U.S.This product is covered
in-part by US 5,871,908 (exp. 12/16/2014) or any foreign equivalents, co-exclusively licensed from Evotec OAI AG. The purchase price includes a license to practice the methods covered by US 5,871,908 (exp. 12/16/2014) by using the product. Purchase of this
product, however, does not convey to the purchaser a license or right to (i) commercially
make, have made or sell reagents and/or kits, or (ii) buy or use reagents and/or kits provided by a third party used in conjunction with the product or any other thermocycler to
practice the methods covered by US 5,871,908 (exp. 12/16/2014) or any foreign equivalents.
RealTime ready
Complex Gene Expression Analysis of Human,
Mouse, and Rat Genes, Made Simple
Maximize efficiency
Select assays for your targets of choice from gene lists of
functionally related gene families or biological pathways. Order
them as single assays or configure your own custom panel on a
LightCycler® 480 Multiwell Plate.
Ensure optimal results
Rely on function tested qPCR assays based on proven Universal
ProbeLibrary technology, with detailed bioinformatic background
information and novel error-detection features.
Get the complete picture
Discover the full functionality of your targets in complex
biological pathways, and reveal their influence on processes
leading to diseases or disorders.
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RealTime ready qPCR Assays and Panels
Get a Head Start on Your Gene Expression Analysis
RealTime ready RT-qPCR Assays for any human,
mouse, or rate gene of your choice are based on
highly specific, LNA-substituted short hydrolysis
probes from the Universal ProbeLibrary (UPL).
Rigorously tested on the LightCycler® 480 Instrument with species-specific universal biological samples, each assay is experimentally qualified to perform according to strict criteria derived from the
MIQE* guidelines.
Linear dynamic range of at least 3 logs
PCR efficiency values of 90% to 110%
R² value of standard curve between 0.99
and 1.00
High amplification specificity – single band in
agarose gel analysis
RealTime ready Assays, like all UPL assays, can
be used successfully on any real-time PCR system
under standard PCR conditions.
Linearity plot (Cp vs log concentration) over five 10-fold dilution
steps for 12 assays from the Human Apoptosis Pathway.
Gel analysis of 48 RealTime ready Assays of the Human GPCR
gene family.
Dilution series of 50 ng to 5 pg of the Human RAN/RANPI
gene, analyzed in triplicates.
* Minimum Information for Publication of Quantitative Real-Time
PCR Experiments
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Assay Design Characteristics
Assay Details
RealTime ready Assays are designed using the proprietary ProbeFinder Software, which is based on
optimized and verified algorithms. For a particular
gene, one representative transcript is chosen for
assay design. The selection of this transcript is
based on annotation and cross-references within
two of the main public gene/transcript annotation
resources, Ensembl and Entrez Gene. This procedure ensures comprehensive coverage of most
known splice variants of a certain gene.
Most RealTime ready Assays are intron-spanning,
enabling reliable quantification unbiased by
genomic DNA residues in the sample material.
For each RealTime ready Assay, the following information is provided:
Accession ID of the transcript for which the assay
was designed
Location of the assay on the selected transcript,
as well as on all other splice variants that were
mapped to the assay by in silico prediction
Amplicon length
Intron/exon boundaries for intron-spanning
assays
Primer sequence and UPL probe number
(provided upon assay delivery)
Chromosomal position
Symbol
Assay ID
Main transcript used for design
Start/end in main transcript
Intron
Exon
3'UTR
Intron-spanning
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RealTime ready Custom Panels for the
LightCycler® 480 System
Custom panels on LightCycler® 480 Multiwell
Plates 96 or 384 are composed of RealTime ready
Assays of your choice. A multitude of different
layouts (see Figures 1 and 2) is available for con­
figuration. Assays are pre-plated and dried down
for a reaction volume of 20 µl on 96-well plates and
10 µl on 384-well plates; simply add sample cDNA
and LightCycler® 480 Probes Master for fast and
reliable results.
Figure 1: Example RealTime ready Custom Panel 96-16+.
On each panel, up to three wells are reserved for
reference gene assays of your choice, matching the
organism you have selected for your assays. In
addition, most panel layouts feature pre-plated
error detection for the respective organism.
Visit www.realtimeready.roche.com to view a short
video demonstration and access detailed information
on how to easily customize your panels. In addition,
you can easily start browsing the Configurator at
www.configurator.realtimeready.roche.com
Figure 2: Example RealTime ready Custom Panel 384-64+.
Error Detection Feature
1 Reference Genes are mandatory controls on each
custom panel that need to be selected during the
custom panel configuration process.
2 RT-Positive Control checks for quality of initial
RNA and the efficiency of the RT step. It consists of
three assays, each targeting different positions of the
same transcript: 3’ end, center region, and 5’ end.
3 RT-Minus Control detects residual genomic DNA.
It consist of two identical assays in two wells: the first
well is for the cDNA sample and the second well is for
the RNA sample.
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RealTime ready Configurator
The RealTime ready Configurator online design tool features an
intuitive graphical interface and a large variety of layouts (with or
without controls) that can be filled in with assays of your choice.
Your configuration progress is automatically stored in your passwordprotected account at every step; your panel content will be available
for download after your purchase is completed.
Browse the Configurator at www.configurator.realtimeready.roche.com.
Search Functions
Assay selection and configuration of custom panels is easy with the
comprehensive search functions of the RealTime ready Configurator.
Search by Pathway: Select assays for your targets from interactive
maps of cellular or signaling pathways.
Search by Focus Panel: Select assays for your targets from preselected lists of large gene families or functionally related groups.
Search by Keyword: Search by gene name, gene symbol, public
accession IDs, biological process, protein function, or a combination of up to 5 keywords. Use Batch Upload to search for up to 384
different accession IDs.
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Search by Focus Panel
RealTime ready Focus Panels
The RealTime ready Configurator features Focus Panels with pre-selected assays for genes from a
variety of pathways, functional groups, and gene superfamilies. The table below shows a selection of
Focus Panels, grouped into different research topics and biological pathways.
Cell Death
Cell Cycle
Development and
Differentiation
Drugable Genome
Angiogenesis
Cell Cycle Checkpoints
Angiogenesis
ABC Transporter
Apoptosis
Cyclin E Lifecycle
Inhibition of Angiogenesis
GPCR Signaling
Caspase Activation
G1/S Transition
Apoptosis
Ion Channels - Ligand Gated
Execution of Apoptosis
M-Phase
Cell Cycle
Ion Channels - Voltage Gated
Induction of Apoptosis
Protein Degradation during Cell
Cycle
DNA Damage and Repair
JNK Signaling
Negative Regulation of Apoptosis
S-Phase
EGF Signaling
Nuclear Receptors
Positive Regulation of Apoptosis
Mitotic Cell Cycle
Downregulation of EGF Signaling
Proteases
Apoptosis - Selection 384
Cell Cycle - Selection 96
Translation Initiation
Protein Kinases
Apoptosis - Selection 96
Apoptosis
GPCRs
Protein Kinases - AGC Group
Proteolytic Processes During
Apoptotic Cell Death
Chromatin Modification
Innate Immunity
Protein Kinases - Atypical Group
DNA Cleavage During Apoptotic
Cell Death
Eukaryotic Translation Initiation
Lipases/Esterases
Protein Kinases - CAMK Group
Cell Cycle
Innate Immunity
PDGF Signaling
Protein Kinases - CK1 Group
Cytochrome P450 Structures
Integrin Family
Proteases
Protein Kinases - CMGC Group
Hemostasis
Neurotrophin Signaling
Protein Kinases
Protein Kinases - Other Group
Integrin Family
Nuclear Receptors
Stem Cell Research
Protein Kinases - RGC Group
PDGF Signaling
Proteases
Generation of IPSCs
Protein Kinases - STE Group
Proteases
Protein Kinases
Hematopoietic Lineage Marker
Genes
Protein Kinases - TK Group
Ectoderm Marker Genes
Protein Kinases - TKL Group
Endoderm Marker Genes
Transcription Factors
Transcription Factors
Mesoderm Marker Genes
Cardio Panel
Small GTPase Mediated Signal
Transduction
Steroid Hormone Receptor
Signaling Pathway
TNFR
Transcription Factors
Tyrosine Kinase Receptor
Signaling
VEGF Signaling
Cardiac Cell Characterization
Smooth Muscle Marker
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Use the Configurator to browse the contents of each of the gene lists below, and
use the “Search by Focus Panel” function to create your own custom panel with
function tested assays. You can either load a complete gene list to your custom
panel or create your own selection by picking the targets from one or more
Focus Panels.
Immune Response
Oncology
Signal Transduction
Apoptosis
Angiogenesis
ABC Transporter
FLT3 Signaling
Apoptosis
Angiogenesis
GPCR Signaling
Cell Cycle
Apoptosis
Hemostasis
Cytochrome P450 Structures
EGF Signaling
Inhibition of Angiogenesis
DNA Damage and Repair
Downregulation of EGF Signaling
Immune System
EGF Signaling
FLT3 Signaling
Innate Immunity
Innate Immunity
GPCR Signaling
Integrins
Integrin Family
Interleukin Signaling
Interleukin Family
Ion Channels
JNK Signaling
Interleukin Receptors
Interleukin Signaling
Neurotrophin Signaling
JNK Signaling
Neurotrophin Signaling
PDGF Signaling
NF-KB Signaling
Nuclear Receptors
RHO GTPase Signaling
NF-KB Target Genes
NF-KB Signaling
SH2 Domain Proteins
Signaling in Immune System
PDGF Signaling
Signaling in Immune System
SH2 Domain Proteins
Proteases
Small GTPase Mediated Signal
Transduction
Hematopoietic Lineage Marker
Genes
Protein Kinases
Steroid Hormone Receptor
Signaling Pathway
TNFR
RHO GTPase Signaling
Translation Initiation
Translation Initiation
Signaling in Immune System
Tyrosine Kinase Receptor
Signaling
Small GTPase Mediated Signal
Transduction
VEGF Signaling
Steroid Hormone Receptor
Signaling Pathway
WNT Signaling
Stem Cell Research
Transcription Factors
VEGF Signaling
WNT Signaling
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Search by Pathway
The “Search by Pathway” function allows you to select your targets
from a variety of interactive maps of cellular or signaling pathways.
These maps immediately reveal possible interactions of your selected
targets with other genes. Simply click on the genes of interest to add
them to your assay search.
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Ordering Information
Product
Cat. No.
RealTime ready Custom Assays
Visit: www.configurator.realtimeready.roche.com
Pack Size
RealTime ready Custom Panels
Visit: www.configurator.realtimeready.roche.com
RealTime ready Focus Panels
RealTime ready Human Reference Gene
Panel, 96
05 339 545 001
2 plates
(each containing 96 assays)
RealTime ready Human Reference Gene
Panel, 384
05 467 675 001
2 plates
(each containing 384 assays)
RealTime ready Human Apoptosis Panel,
96
05 392 063 001
2 plates
(each containing 96 assays)
RealTime ready Human Apoptosis Panel,
384
05 339 316 001
2 plates
(each containing 384 assays)
RealTime ready Human ABC
Transporter Panel, 96
05 339 324 001
2 plates
(each containing 96 assays)
RealTime ready Human ABC
Transporter Panel, 384
05 467 713 001
2 plates
(each containing 384 assays)
RealTime ready Human Cell Cycle
Regulation Panel, 96
05 339 359 001
2 plates
(each containing 96 assays)
RealTime ready Human Cell Cycle
Regulation Panel, 384
05 467 683 001
2 plates
(each containing 384 assays)
RealTime ready Human GPCR Panel, 96
05 353 068 001
2 plates
(each containing 96 assays)
RealTime ready Human GPCR Panel, 384
05 467 705 001
2 plates
(each containing 384 assays)
RealTime ready Human Nuclear Receptor
Panel, 96
05 339 332 001
2 plates
(each containing 96 assays)
RealTime ready Human Nuclear Receptor
Panel, 384
05 467 691 001
2 plates
(each containing 384 assays)
Real-Time PCR Reaction Mixes
RealTime ready DNA Probes Master
05 502 381 001
5 x 1 ml (5x conc.)
LightCycler® 480 Probes Master
04 707 494 001
5 x 1 ml (2x conc.)
RealTime ready Cell Lysis Kit
05 943 523 001
1 Kit (500 reactions)
Transcriptor First Strand cDNA Synthesis
Kit
04 379 012 001
1 Kit (50 reactions)
Transcriptor Universal cDNA Master
05 893 151 001
100 reactions
LightCycler 480 Sealing Foil
04 729 757 001
50 foils
Additional Products
®
Roche Applied Science offers a large selection of
reagents and systems for life science research. For a
complete overview of related products and manuals,
visit and bookmark our homepage,
www.roche-applied-science.com, and our Special
Interest Sites including:
RealTime ready Custom Assays and Panels:
http://www.realtimeready.roche.com
The Universal ProbeLibrary:
http://www.universalprobelibrary.com
The LightCycler® System family for real-time PCR:
http://www.lightcycler.com
Amplification – Innovative tools for PCR:
http://www.roche-applied-science.com/pcr
Trademarks:
LIGHTCYCLER, REALTIME READY, and NIMBLEGEN are trademarks of Roche.
Exiqon, LNA ProbeFinder and ProbeLibrary are registered trademarks of Exiqon
A/S, Vedbaek, Denmark.
For life science research only.
Not for use in diagnostic procedures.
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Published by
Roche Diagnostics GmbH
Sandhofer Straße 116
68305 Mannheim
Germany
© 2011 Roche Diagnostics. All rights reserved.
www.roche-applied-science.com
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