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LightCycler® Multiplex DNA Master
LightCycler® Multiplex RNA Virus Master
Made to discover more
Introduction
Introducing the new
LightCycler® Masters
optimized for Multiplex qPCR
5x advantage
The initial steps of your experiments set an important foundation to your
work. Hence it’s important to get the details right in order to ensure
successes down the line.
Bring your qPCR to the next level with two newly formulated master mixes
from Roche that can detect a greater number of targets per reaction, work
more efficiently, and maximize your valuable samples.
Robust results
Select one of the new master mixes
for efficient and reliable detection of:
Improve workflow
• R
NA targets (one-step) –
LightCycler®Multiplex
RNA Virus Master
• D
NA targets (two-step) –
LightCycler® Multiplex
DNA Master
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• More freedom to include multiple sets of oligos (probes and primers)
• Less volume taken up by the mix, so more volume is available for the
sample material, providing a more sensitive target detection
Our
5x
concentrated
LightCycler®
Multiplex Masters
• Detect low copy number of DNA or
RNA targets with higher sensitivity
5x advantage
The advantages of a 5x concentrated mix in multiplex qPCR include:
Introduction
5x concentrated to deliver
outstanding performance
and throughput
• Suitable for a wide range
of sample materials
• Increase your throughput
and work efficiency
• Allow you to achieve maximum data
from your valuable samples
Robust results
Other considerations
when choosing a master
mix for multiplexing
Improve workflow
aster mix developed for successful, reliable multiplex
M
qPCR must be engineered to:
• Achieve high specificity amplicon generation in the
presence of multiple primer pairs
in the amplification mixture
• Ensure the correct targets are amplified, while
minimizing non-specific primer artifacts
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Analyze your results with confidence
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• Be robust enough to drive several amplifications
per reaction
Introduction
For DNA targets
The LightCycler® Multiplex RNA Virus Master
The LightCycler® Multiplex DNA Master
• Optimized for fast and reliable detection
of up to five RNA targets
• Optimized for fast and reliable detection
up to five DNA targets
• Consists of a unique recombinant reverse transcriptase
and the aptamer-mediated hot start AptaTaq
Fast DNA Polymerase
• Specially formulated for fast and highly specific multiplex
qPCR or two-step RT-qPCR
• Guarantees fast and highly specific one-step multiplex
qPCR detection of RNA targets isolated from a variety of
sample materials
• Guarantees fast and highly
specific multiplex qPCR
detection of DNA targets
isolated from a variety of
sample materials
Improve workflow
• Ensures RNA test
results are derived
from RNA transcripts,
not from residual
genomic DNA through
a two-vial composition
separating RT enzyme
from RT minus controls
Robust results
For RNA targets
• Guarantees fast and highly specific one-step multiplex
RT-qPCR detection of RNA targets isolated from a variety
of sample materials
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5x advantage
Rely on the new LightCycler®
Multiplex Masters to maximize
the performance of your
LightCycler® System
β-Globin (FAM)
Similar amplification curves, in
singleplex vs. 4-plex reactions
C
Similar amplification curves, in
singleplex vs. 4-plex reactions
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your valuable samples
5
10
15
20 25
Cycles
30
35
40
45
5
10
15
20 25
Cycles
30
35
40
45
5
10
15
20 25
Cycles
30
35
40
45
5
10
15
20 25
Cycles
30
35
40
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• Rely on a master mix optimized for multiplex detection of up to five targets
• Detect low copy numbers of DNA or RNA targets with higher sensitivity
using a 5x master mix concentration
5x advantage
Increase throughput and cost efficiency of your qPCR
Introduction
C
Parvo B19 (HEX)
• Expect equivalent performance in multiplex or singleplex
qPCR reactions (Fig. 1)
PBGD (Red610)
Average Cq Values
β-Globin Linearity
A
B
Comparable Cq Values, in singleplex vs.
4-plex reactions
C
Control (Cy5)
Similar amplification curves, in
singleplex vs. 4-plex reactions
C
Similar amplification curves, in
singleplex vs. 4-plex reactions
Same linearity, in singleplex vs.
4-plex reactions
Robust results
• Save cost and time by making your samples go further
40
40
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32
30
>
30
5
10
15
20
25
30
35
40
45
5
10
15
20 25
Cycles
30
35
40
45
30
35
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5
10
15
20 25
Cycles
30
35
40
45
Cycles
28
26
25
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22
20
ß-globin
Singleplex
PBGD
Control
2.50
-2.00
4-plex
-1.50
Singleplex
-1.00
-0.50
20
-0.00
5
10
15
20 25
Cycles
Figure 1. Results show similar Cq values (A), PCR efficiency (B) and amplification curves (C)
of 4 different targets, when ran in singleplex and 4-plex reactions. Four different
assays were ran on a LightCycler® 96 System, using the LightCycler® Multiplex DNA Master.
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4-plex
Parvo B19
Introduction
Generate consistent results
β2M and Control Singleplex and Duplex
Reproducible Multiplex qPCR, from one person to another
Achieve robust and reproducible results,
while minimizing optimization
Operator 1
Operator 2
57.490
52.490
47.490
42.490
37.490
32.490
27.490
22.490
17.490
12.490
7.490
2.490
• Obtain linear, sensitive and reproducible results in a
multiplex qPCR (Fig. 2)
• Deliver consistent and reproducible multiplex performance, from one
colleague to another (Fig. 3)
• Include reference gene assays, such as Universal ProbeLibrary (UPL)
assays from Roche, in the same reaction as your targets
0
45.707
41.707
36.707
31.707
26.707
21.707
16.707
11.707
6.707
1.707
5
10
Robust performance across a wide dynamic range
Robust performance across a wide dynamic range
β-Globin (FAM-labeled)
β2M (Yellow 555-labeled)
27.571
44.617
Fluorescence (533-580)
49.617
24.571
21.571
18.571
15.571
12.571
9.571
6.571
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15
20
25
Cycles
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35
40
45
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40
0
45
2
6
10
14
18 22 26
30 34 38
42
Cycles
NTC
Singleplex
Duplex
NTC
Reproducible Multiplex qPCR, from one person to another
Operator 1
Operator 2
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40
35
35
34.617
30
30
29.617
25
25
24.617
20
20
15
15
10
10
5
5
39.617
19.617
14.617
0
-4.00 -3.50 -3.00 -2.50 -2.00 -1.50 -1.00 -0.50 0.00 -4.50
0.383
5
30
β2M Linearity
4.617
0.571
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10
15
20
25
30
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40
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Cycles
β2M Singleplex
β2M Duplex
-5.00
-4.00
β2M Singleplex
-3.00
-2.00
-1.00
0
0.00
β2M Duplex
Figure 3. Results show similar Cq values, qPCR efficiency and sensitivity of β2M,
in triplicate, singleplex and duplex reactions, even when performed by different
operators. β2M gene was detected with the LightCycler® 480 System, using the
LightCycler® Multiplex DNA Master.
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Figure 2. Expect the same robust performance across a dynamic range. Duplex assays of serial
1:4 dilutions (10 ng to 2.4 pg) on the LightCycler® 480 System, using the LightCycler® Multiplex
DNA Master, were detected with a β-globin UPL (Universal ProbeLibrary) probe and a UPL
reference gene assay for β2M.
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Duplex
9.617
3.571
25
Improve workflow
Fluorescence (465-510)
β2M (Yellow 555-labeled)
30.571
20
Robust results
β-Globin FAM-labeled
54.617
15
Cycles
Singleplex
33.571
5x advantage
• Detect a multitude of targets without the need for reaction optimization
51.707
• Decrease experiment set-up time and effort with a convenient
single vial mix for DNA
With many labs having to work with finite resources, multiplex qPCR
tests are a great way for your lab to run more efficiently and realize
significant cost savings.
• Ensure RNA test results are derived from RNA transcripts, not from
residual genomic DNA through a two-vial composition separating RT
enzyme from RT minus controls
• Experience peace of mind when setting up room temperature reactions
with these new thermostable masters
• Increase your lab throughput with mixes compatible with automation
Multiplex qPCR detection offers several advantages. It can increase your
efficiency, maximize your valuable samples, and allow you to easily include
reference gene with your targets.
5x advantage
Designed to fit the growing
needs of your lab
Introduction
Improve your qPCR workflow
Now that you‘ve chosen a LightCycler® system, make the most of your qPCR
research and rely on the experienced technical team at Roche for both your
instrument and application support.
Robust results
Improve workflow
Make a difference to the way
you discover more with the
LightCycler® Multiplex Masters
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Introduction
Order today and start
to discover more
Cat No
No. of rxns
Content
5x advantage
LightCycler® Multiplex RNA Virus Master
06 754 155 001
07 083 173 001
200 reactions of 20µl
1000 reactions of 20µl
1. RT-Enzyme Solutions,
200x conc.
2. RT-qPCR Reaction Mix,
5x conc.
3. Water, PCR grade
LightCycler® Multiplex DNA Master
200 reactions of 20µl
07 339 577 001
1000 reactions of 20µl
Robust results
07 339 585 001
1. qPCR Reaction Mix, 5x conc.
2. Water, PCR grade
Improve workflow
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Learn more about the LightCycler® Multiplex Masters at
lifescience.roche.com/Multiplex
Published by
Roche Molecular Systems, Inc.
4300 Hacienda Drive,
Pleasanton, CA 94588
©2015 Roche Molecular Systems Inc.
For general laboratory use.
LIGHTCYCLER is a registered trademark of Roche.