1. The Most Reliable SYBR® Green-based Real-Time qPCR Assays for Gene Expression Analysis
RT² qPCR Primer Assays are the most reliable gene expression
analysis tool using SYBR Green-based quantitative real-time PCR
Verified Specificity
Ct = 27.3
Ct = 21.1
technology. Each assay utilizes a proprietary and experimentally
verified algorithm for designing gene-specific qPCR primers
with uniform PCR efficiency and amplification conditions. Every
RT² qPCR Primer Assay is subjected to rigorous experimental
verification. Amplification of a single product of the correct
size and high PCR efficiency (>90%) are guaranteed when the
assays are used with the RT² qPCR Master Mixes. With high
PCR efficiency and uniform PCR conditions, it is convenient to
perform quantitative real-time PCR for several genes under the
MMP13
MMP15
Each RT² qPCR Primer Assay is experimentally verified to yield a single
dissociation curve peak and to generate a single amplicon of the correct
size for the targeted gene.
Wide Linear Dynamic Range
same PCR conditions.
Benefits of RT2 qPCR Primer Assays
High Performance:
Each RT² qPCR Primer Assay is experimentally
validated to amplify a single amplicon of correct size at
uniform PCR efficiency. The performance of RT² qPCR
Primer Assays is similar to that of TaqMan® assays.
Complete Genome Coverage:
RT² qPCR Primer Assays are available for every human,
mouse and rat gene. Their uniform PCR efficiency and
PCR conditions allows easy transition from single gene
analysis to multiple gene expression profiling.
Convenience:
With less than 5-minutes of hands-on time, RT² qPCR
Primer Assays deliver guaranteed performance
when used with optimized RT² qPCR Master Mixes.
Master Mixes are available for all real-time PCR
instruments.
Wide linear dynamic range enables simultaneous analyses for rare and
abundant transcripts. Ten-fold serial dilutions of purified DNA were used
as template in RT² qPCR Primer Assays. The qPCR was run in duplicate
for each dilution point. The RT² qPCR Primer Assays detect from 1 copy
to 1010 copies of template covering a linear dynamic range of up to eight
(8) orders of magnitude.
Equivalent Performance to TaqMan Assays
TaqMan Gene Expression Assays
Fold Change (log2)
qPCR ASSAYS
RT2 qPCR Primer Assays
20
15
10
5
-20
-15
-10
-5
5
10
15
20
-5
-10
-15
-20
RT2 qPCR Primer Assays
Fold Change (log2)
A high concordance is observed between RT² qPCR Primer Assays
and TaqMan Gene Expression Assays. Gene expression comparisons
were made between two samples for 84 genes by the RT² qPCR Primer
Assay method and the TaqMan Gene Expression Assay method. Fold
changes (log 2 ) in expression for each gene are calculated and plotted
against one other for the two methods. The gray dashed line represents a straight line with an ideal slope of 1.0. The solid black line
shows the linear regression data fit. The Pearson correlation coefficient (R value ) between the two sets of data is 0.97.

2. Why Use RT² qPCR Primer Assays?
RT² qPCR Primer Assay Features
SYBR® Green assays do not require the design of a specific probe
High Specificity
Single band of correct size
in addition to the primers for each gene-specific assay, reducing
Uniform Efficiency
>90% PCR efficiency*
the assay setup time and running costs. A drawback of SYBR
High Sensitivity
As low as 10 copies per cell
Green assays, however, is that the dye binds all double-stranded
Wide Dynamic Linear Range
10 - 109 copies
DNA non-specifically and can generate false positive signals
if non-specific products and primer dimers are present in the
assay. Conventional PCR primers, not specifically designed for
Applications
real-time PCR, often have the problems of low PCR amplification
efficiency, primer dimers, and non-specific amplification. In
Validate microarray-derived gene expression data.
short, conventional PCR primers can not be used for real-time
PCR.
Examine a focused panel of genes using multiple
RT² qPCR Primer Assays.
RT² qPCR Primer Assays are specifically designed for real-time
PCR analysis. The assay design criteria ensure that each qPCR
Verify gene expression knock down by RNAi.
reaction will generate single, gene-specific amplicons and prevent
the co-amplification of non-specific products. Furthermore,
Identify and confirm disease-associated
simultaneous gene expression analyses require similar PCR
biomarkers.
efficiencies for accurate comparisons among genes. RT² qPCR
Monitor phenotypic changes related to gene
Primer Assays are designed with an amplicon size range of 100-
expression.
250 bp and their PCR efficiency* is uniformly >90%. Overall,
more than 10 thermodynamic criteria are included in each RT²
qPCR Primer Assay design to deliver the most reliable and
* PCR Efficiency is calculated using DART-PCR workbook
(Nucleic Acid Research 2003 31 (14): e73.)
accurate gene expression analysis results.
RT2 qPCR Primer Assays Ordering Guide
www.SABiosciences.com/RT2PCR.php
Find the qPCR Assay for your genes at:
Description
Gene-Specific RT2 qPCR Primer Assay
Cat. No.
PPX#####Y-200
X = H, Human; M, Mouse; R, Rat
#####Y identifies specific gene and design
For Guaranteed qPCR Performance, Use RT2 Reagents:
RT2 qPCR Master Mixes
Instrument Type
Cat. No.
RT2 SYBR Green / ROX qPCR Master Mix
Applied Biosystems, Stratagene, Roche
PA-112 (2000 qPCR reactions)
2
RT SYBR Green / Fluorescein qPCR Master Mix Bio-Rad
PA-111 (2000 qPCR reactions)
RT2 SYBR Green qPCR Master Mix
PA-110 (2000 qPCR reactions)
RT2 First Strand Kit
C-03
RT2 qPCR-Grade RNA Isolation Kit
PA-001 (12 RNA isolations)
All other real-time instruments
(12 first strand reactions)
RT2 qPCR Primer Assays™ and PCRGrade™ are trademarks of SABiosciences Corporation. SYBR® is a registered trademark of Molecular Probes, Inc. ABI® and ROX® are a registered
trademarks of Applera Corporation. Bio-Rad® is a registered trademark of Bio-Rad Laboratories. Stratagene® is a registered trademarks of Stratagene. TaqMan® is a registered trademark of Roche
Molecular Systems.
SABiosciences Corporation
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Frederick, MD 21703
www.SABiosciences.com
T 301.682.9200
888.503.3187
F 301.682.7300
888.465.9859