Amplification: PCR Reagents

iQ Multiplex Powermix

iQ™ Multiplex Powermix

iQ multiplex powermix simplifies real-time detection of multiple targets in a

single tube, routinely yielding efficiencies equivalent to corresponding singleplex
reactions. Increase throughput, control costs, and maximize your data output
with this powerful blend.

Reliable real-time multiplex PCR

n 

detection of up to 5 targets
n Detection of up to 4 targets, one of
which may differ 106-fold in expression
n Linearity over 6 orders of magnitude of
input cDNA and 4 orders of magnitude
of input genomic DNA

For more information, visit us on the Web

at www.bio-rad.com/supermixes/
 FAM: a-tubulin

1,000
Detect Multiple Targets Without Optimization
Bio-Rad has applied its expertise in multiplex

PCR baseline-subtracted RFU

real-time PCR to create a robust mix that 1,000
100
greatly simplifies real-time detection of
multiple targets in a single tube. Although 100
0 2 6 10 14 18 22 26 30 34 38 42 46 50
design software, such as Beacon Designer FAM: a-tubulin HEX: a-tubulin
FAM: GAPDH
software, has made it easier to design effective 1,000
35 1,000
1,000 0 5 10 15 20 25 30 40
primers and probes, finding a set of reaction Cycle
conditions that amplifies all targets with equal Comparable sensitivity between four-plex assay and corresponding singleplex assays
efficiency in both singleplex and multiplex on the iQ™5 real-time PCR detection system. Spleen cDNA amplified with primers to 18S

PCR base line subtracted RFU PCR base line subtracted RFU
rRNA (HEX-labeled probe), b-actin (FAM-labeled probe), tubulin (Cy5-labeled probe), and IL-2
reactions can still be a challenge. Until now, 100
(Texas 100or multiplex (green traces). There was
100
Red-labeled probe) in either singleplex (red traces)
optimization entailed an empirical approach no loss in sensitivity on converting these four single assays into a multiplex assay, as seen by the
in which buffer components, enzyme close threshold
0 2 6 crossings
10 14 18 of22singleplex
26 30 34vs. 42 46 50traces.0022 66 10
38multiplex 10 14
14 18
18 22
22 26
26 30
30 34
34 38
38 42
42 46
46 50
50

concentrations, dNTP amounts, and primer a-Tubulin

FAM: a-tubulin
HEX: GAPDH TexasGAPDH
GAPDH
HEX: Red: IL–1b

concentrations were adjusted to ensure similar 1,000

1,000 1,000
1,000
amplification curves in singleplex and multiplex
reactions. Consequently, many researchers
have avoided performing multiplex real-time RFU
line subtractedRFU
PCR or only perform this time-consuming 100
100 100
100
baseline-subtracted

process when it is absolutely required for

0 2
0 2 6 10 14
6 10 14 18
18 22
22 26
26 30
30 34
34 38
38 42
42 46
46 50
50 0 2
0 2 66 10
10 14
14 18
18 22
22 26
26 30
30 34
34 38
38 42
42 46
46 50
their experiment.
IL-1b
HEX: GAPDH
Texas Red: IL–1b Cy5: factor
Factor
Texas Red: VIII
VIII IL–1b
To help simplify multiplex real-time PCR,
1,000 1,000
1,000
base

1,000 1,000
Bio-Rad has developed iQ multiplex powermix.
PCR

This mix makes multiplex real-time PCR easier

PCR base line subtracted RFU PCR

by eliminating the need to optimize buffer,

enzyme, or primer concentrations.
100
100 100
100
100

Save time and reduce reagent costs —

000 2
22 66
0 2
0 10
6 10
2 6 14
10 14 18
14 18
10 14
6 10 22
18 22
14 18 26
22 26
18 22 30
26 30 34
30 34
22 26 38
34 38
26 30 42
38 42
30 34 46
42 46
34 38 50
46 50
38 42
42 46
46 50
50
with this reliable mix, you can increase Cycle Cycle
Cycle
throughput and control costs by running Texas Red: IL–1b
Cy5: factor VIII Cy5: factor VIII
Linearity of four-target detection using the iQ5 real-time PCR detection system. A series
multiple assays in a single reaction, of 10-fold
1,000 dilutions of human genomic DNA (500 ng–50 pg in a 50 µl reaction) was amplified to
1,000
assay for four target genes using iQ multiplex powermix. Targets were a-tubulin, detected with
maximizing the amount of data collected 2
a FAM-labeled probe (efficiency = 96.4%, r = 0.996); GAPDH, detected with a HEX-labeled
from limited amounts of sample. iQ multiplex probe (efficiency = 94.9%, r2 = 0.998); IL-1b, detected with a Texas Red-labeled probe
powermix is formulated for analysis using (efficiency = 101.8%, r2 = 0.998); and factor VIII, detected with a Cy5-labeled probe
(efficiency = 101.6%, r2 = 0.994).
cDNA, genomic DNA, and plasmids, 100 100
and can be used for a wide variety of
0 2 6 10 14 18 22 26 30 34 38 42 46 50 0 2 6 10 14 18 22 26 30 34 38 42 46 50
applications, including: Ordering Information Cycle
Cycle
Cy5:
Catalog # factor VIII
Description
n Gene expression analysis
172-5848 
1,000 iQ Multiplex Powermix, 50 x 50 µl reactions, 2x mix contains dNTPs,
n SNP genotyping/SNP analysis 11 mM MgCl2, iTaq™ DNA polymerase, stabilizers
172-5849 iQ Multiplex Powermix, 200 x 50 µl reactions, 2x mix contains dNTPs,
n GMO detection 11 mM MgCl2, iTaq DNA polymerase, stabilizers
n Viral load detection
Beacon Designer is a trademark of PREMIER Biosoft International. Cy is a trademark of GE Healthcare group companies.
Texas100
Red is a trademark of Invitrogen Corporation.
Practice of the patented 5' Nuclease Process requires a license from Applied Biosystems. The purchase of this product
includes an immunity from suit under patents specified in the product insert to use only the amount purchased for the
purchaser’s0own2 internal
6 10research
14 18when22 used
26 30with34 38 42 purchase
the separate 46 50 of Licensed Probe. No other patent rights are
conveyed expressly, by implication, orCycle
by estoppel. 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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Bulletin 5348 US/EG Rev B 07-0855 1207 Sig 1207