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Got Limited Sample?
PreAmp It!
PCR Reagents: SsoAdvanced™ PreAmp Supermix and PrimePCR™ PreAmp Assays
Bulletin 0000
6576
What is PreAmp?
PreAmp provides the ultimate in unbiased, 1,000-fold target-specific preamplification from
limited nucleic acids, using up to 100 quantitative PCR (qPCR) or end-point assays and
enabling more downstream reactions for screening of respective targets.
Why use SsoAdvanced PreAmp Supermix?
■■ Unrivaled, with half the bias specification of the competition
■■ Requires as little as 100 pg of genomic DNA or cDNA
■■ Validated for SYBR® Green and probe chemistries
■■ Optimized for PrimePCR PreAmp Assays
■■ Improved performance using TaqMan and custom assays
■■ Superior sensitivity and efficiency
■■ Economically priced
Why use PrimePCR PreAmp Assays?
■■ Available for each PrimePCR gene expression assay
■■ SYBR® Green and probe versions offered
■■ Easy to use — no dilutions needed
■■ Proven performance
■■
■■
ull wet-lab validation ensures assays meet the minimum
F
information for publication of quantitative real-time PCR
experiments (MIQE) guidelines
Optimized for use with SsoAdvanced PreAmp Supermix
What types of samples work with PreAmp?
■■ Plant samples
■■ Formalin-fixed, paraffin-embedded samples
■■ Stem cells
■■ Single cells
■■ Small biopsies
■■ Laser capture microdissection samples
■■ Rare samples
Visit bio-rad.com/web/PreAmp1 for more information.
SsoAdvanced™ PreAmp Supermix and PrimePCR™ PreAmp Assays
How do PreAmp products compare to others?
A
Bio-Rad
D
3.00
Twenty-four qPCR probe assays with their
respective qPCR reagents were used to
evaluate the level of bias.
0.75
–0.75
5
10
15
Number of qPCR assays
–3.00
0
25
Bio-Rad
Eighty-eight qPCR SYBR® Green assays with
their respective qPCR reagents were used
to evaluate the level of bias.
PrimePCR™ PreAmp SYBR ® Green Assays,
SsoAdvanced PreAmp Supermix, and
SsoAdvanced™ Universal SYBR® Green Supermix
RT 2 Assays, RT 2 qPCR Primer Assay, RT2 PreAMP
Pathway Primer Mix, and RT2 qPCR Master Mix
Bio-Rad
®
Ninety qPCR SYBR Green assays with
their respective qPCR reagents were used
to evaluate the level of bias.
2.25
1.50
0.75
PrimePCR™ PreAmp SYBR® Green Assays,
SsoAdvanced PreAmp Supermix, and
SsoAdvanced™ Universal SYBR® Green Supermix
0.00
–0.75
–1.50
PerfeCTa PreAmp 5x SuperMix and
PerfeCTa® SYBR® Green FastMix
–2.25
22
26
Cq value
10
15
Number of qPCR assays
30
20
25
QIAGEN
3.00
2.25
1.50
0.75
0.00
–0.75
–1.50
–2.25
–3.00
–3.75
–4.50
–5.25
–6.00
–6.75
–7.50
22
24
26
F
3.00
–3.00
18
5
E
3.00
2.25
1.50
0.75
0.00
–0.75
–1.50
–2.25
–3.00
–3.75
–4.50
–5.25
–6.00
–6.75
–7.50
16 18202224 2628303234
Cq value
C
–0.75
–2.25
BQ score
B
20
0.75
0.00
–1.50
TaqMan Gene Expression Assays, TaqMan
PreAmp Master Mix, and TaqMan Gene
Expression Master Mix
–2.25
BQ score
1.50
PrimePCR PreAmp Probe Assays, SsoAdvanced
PreAmp Supermix, and SsoAdvanced Universal
Probes Supermix
0.00
–1.50
BQ score
2.25
BQ score
BQ score
1.50
BQ score
2.25
–3.00
0
Life Technologies
3.00
28
30
Cq value
32
34
Quanta BioSciences
2.25
1.50
0.75
0.00
–0.75
–1.50
–2.25
–3.00
–3.75
–4.50
–5.25
–6.00
–6.75
–7.50
22
24
26
28
Cq value
30
32
34
What is a BQ score?
A bias quantification (BQ) score provides a numerical value for the level of bias observed in a PreAmp reaction. It is based
on comparing the theoretical difference between a sample with and one without PreAmp compared to the observed (actual)
difference between a sample with and without PreAmp. Zero indicates no bias, and the farther from zero the BQ score
becomes, the greater the bias introduced.
Want more details?
Property
SsoAdvanced
PreAmp Supermix
TaqMan PreAmp Master Mix
(Life Technologies)
PerfeCTa PreAmp 5x SuperMix
(Quanta BioSciences)
BQ score
90% ≤ 1.50
90% ≤ 0.75
90% ≤ 1.50
Sensitive detection of low-level target genes
•••
••
•
High efficiency, even for difficult amplicons
•••
••
••
PrimePCR PreAmp, TaqMan
TaqMan
Unknown
SYBR® Green and probes
Probes
SYBR® Green and probes
50 x 50 µl
40 x 50 µl
40 x 50 µl
Validated assays
Chemistry
Number of reactions
•Fair
••Good
•••Excellent
© 2014 Bio-Rad Laboratories, Inc.
Bulletin 6576
SsoAdvanced™ PreAmp Supermix and PrimePCR™ PreAmp Assays
How does it work?
From RNA to qPCR results, data can be obtained in less than 5 hours using our validated reverse transcription quantitative PCR
(RT-qPCR) products.
1
Extract RNA
2
3
Produce cDNA
Combine components
4
Perform preamplification PCR
cDNA
Reverse transcription with
iScript™ Reverse Transcription Kits
5
Dilute sample
1:5 to 1:20
6
Preamplified
sample
Set up qPCR reaction
Primers
7
10 –12 cycles
SsoAdvanced
PreAmp Supermix
Perform qPCR
SsoAdvanced
Universal
Supermix
8
Analyze data
104
RFU
Pooled
primers
103
102
0 1020 3040
Cycles
Preamplification workflow.
How do I order SsoAdvanced PreAmp Supermix?
Order online at bio-rad.com/web/PreAmpSmx, or contact your local sales representative
to request a quote for SsoAdvanced PreAmp Supermix (catalog #172-5160).
How do I order PrimePCR PreAmp Assays?
Order online at bio-rad.com/web/PrimePCRPreAmp, or contact your local sales representative
for additional information.
What other validated reagents do I need?
iScript cDNA Synthesis Kits provide unbiased representation of target mRNA levels through the use of an RNase H+ Moloney
murine leukemia virus (MMLV) reverse transcriptase and optimal priming methodology using a blend of oligo(dT) and random
primers. iScript kits are available in single- or multiple-tube formats
■■
— Order online at bio-rad.com/web/iScriptKits, or contact your local sales representative for additional information
■■
soAdvanced Universal Supermixes employ our patented* Sso7d fusion enzyme technology for higher processivity,
S
increased PCR inhibitor tolerance, and robust performance with challenging templates and target sequences for all real-time
PCR applications and all real-time PCR systems. SsoAdvanced Universal Supermixes are available in SYBR® Green and
probe versions
— Order online at bio-rad.com/web/SsoAdvanced, or contact your local sales representative for additional information
* U.S. patents 6,627,424; 7,541,170; and 7,560,260.
© 2014 Bio-Rad Laboratories, Inc.
Bulletin 6576
SsoAdvanced™ PreAmp Supermix and PrimePCR™ PreAmp Assays
PerfeCTa is a trademark of Quanta BioSciences, Inc. RT2 is a trademark of SABiosciences Corporation. SYBR is a trademark of
Life Technologies Corporation. Bio-Rad Laboratories, Inc. is licensed by Life Technologies Corporation to sell reagents containing
SYBR Green I for use in real-time PCR, for research purposes only. TaqMan is a trademark of Roche Molecular Systems, Inc.
Bio-Rad’s real-time thermal cyclers are covered by one or more of the following U.S. patents or their foreign counterparts owned by
Eppendorf AG: U.S. Patent Numbers 6,767,512 and 7,074,367.
The use of SsoAdvanced Supermixes and PrimePCR PreAmp Assays is covered by one or more of the following U.S. patents and
corresponding patent claims outside the U.S.: 5,804,375; 5,538,848; 5,723,591; 5,876,930; 5,994,056; 6,030,787; 6,171,785; and
6,258,569. The purchase of these products 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. No right under any other patent claim and no right to
perform commercial services of any kind, including without limitation reporting the results of purchaser’s activities for a fee or
other commercial consideration, are conveyed expressly, by implication, or by estoppel. These products are 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.
Bio-Rad
Laboratories, Inc.
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Life Science
Group
Bulletin 6576 Rev C
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14-1558
0814
Sig 1213