1. Performance of Applied Biosystems™ consumables on Bio-Rad CFX™ Real-Time PCR Detection System

2. Applied Biosystems™ Consumables on Bio-Rad CFX™
Demonstrate performance of Applied Biosystems consumables on the Bio- Rad CFX96 and CFX384 Real-Time PCR detection systems*.
TaqMan® Fast Advanced Master Mix with TaqMan® Gene Expression Assays
Reproducibility
Dynamic Range
Power Up™ SYBR™ Green Master Mix
Specificity
*Bio-Rad, CFX96 and CFX384 are trademarks of Bio-Rad Laboratories, Inc., which is not affiliated with Thermo Fisher. The statements herein have not been reviewed or endorsed by Bio-Rad.

3. Materials TaqMan® Fast Advanced Master Mix
TaqMan® Gene Expression Assays:
18S Hs _s1 VIC Primer-limited
GPR6 Hs _s1 FAM
PPIA Hs _m1 FAM
YWHAZ Hs _g1 FAM
cDNA: Universal Human Reference (UHR)
PowerUp™ SYBR™ Green Master Mix
Targets (all primers run at final concentration of 300nM)
A2M ADAM10
AOF1 APOA1
ARL1 C11orf30
GAPDH PGK1
cDNA: UHR or liver
All reactions run in quadruplicates
CFX96 – 20ul rxns
CFX384 – 10ul rxns

4. TaqMan Fast Advanced Master Mix protocol on Bio-Rad CFX96™ or CFX384™
(Optional UNG step)
Note: Use an anneal temperature appropriate for your assay or primer pair design

5. TaqMan Fast Advanced Master Mix: Reproducibility
From dynamic range data
with 4 replicates
GAPDH
GPR6
PPIA
YWHAZ
At high Ct’s detection is near single copy, therefore precision becomes less reliable.
Cq < 32

6. TaqMan Fast Advanced Master Mix: Six log dynamic range
GAPDH
Dynamic range is likely wider, but didn’t run higher concentrations of cDNA
PPIA
YWHAZ

7. PowerUp™ SYBR™ Green Master Mix
An optional UNG treatment step at 50 ºC may be added at the start of the run.
Use an annealing temperature appropriate for the primer pair.
50 ºC / 2 min (optional UNG step)
95 ºC / 2 min
95 ºC / 3 sec x40 cycles
60 ºC / 30 sec
Melt curve

8. PowerUp™ SYBR™ Green Master Mix: Dynamic Range
Excellent efficiency and reproducibility
Cq < 32

9. PowerUp™ SYBR™ Green Master Mix: PGK1
R2 value of 99.9% shows excellent linearity over a wide input range
Single peak showing high target specificity
Steep curves show high efficiency

10. PowerUp™ SYBR™ Green Master Mix: ARL1
R2 value of 99.8% shows excellent linearity over a wide input range
Low peak (1 pm) is equivalent to the low plateau
Single peak showing high target specificity
Steep curves show high efficiency

11. A2M AOF1 ARL1 ADAM10 APOA1 C11orf30
PowerUp™ SYBR™ Green Master Mix: Excellent Reproducibility, Six targets
A2M
AOF1
ARL1
ADAM10
APOA1
C11orf30

12. PowerUp™ SYBR Green Master Mix: Excellent Specificity – Dissociation Data
APOA1
ARL1
C11orf30
A2M
ADAM10
AOF1
No non-specific amplification or primer dimer

13. Conclusions and Next Steps
Applied Biosystems™ consumables perform well on the Bio-Rad CFX™ (96 & 384 well platforms)
TaqMan® Gene Expression Assays run with TaqMan® Fast Advanced Master Mix shows excellent reproducibility, dynamic range and equivalent results in singleplex vs duplex.
PowerUp™ SYBR™ Green Master Mix shows excellent reproducibility and sensitivity.
To learn more about Applied Biosystems qPCR master mixes or to request a free sample, please visit
To download the ‘Design and Optimization of SYBR Green Assays’ and/or ‘Protocol for Comparing SYBR Green Master Mixes’ please visit
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PowerUp
PGK1 – 4 logs
ARL1 – 4 logs
ABCC2 – 2 logs
© Thermo Fisher Scientific Inc. All rights reserved. All trademarks are the property of Thermo Fisher Scientific and its subsidiaries unless otherwise specified. TaqMan is a registered trademark of Roche Molecular Systems, Inc., used under permission and license. For research use only. Not for use in diagnostic procedures.