Performance of Applied Biosystems consumables on BioRad CFX

Performance of Applied Biosystems™ consumables on Bio-Rad CFX™ Real-Time PCR Detection System 1 The world leader in serving science

Applied Biosystems™ Consumables on Bio-Rad CFX™ Demonstrate performance of Applied Biosystems consumables on the Bio. Rad CFX (96 & 384 well) instruments. • Taq. Man® Fast Advanced Master Mix with Taq. Man® Gene Expression Assays • Reproducibility • Dynamic Range • Power Up™ SYBR™ Green Master Mix • Reproducibility • Specificity • Dynamic Range 2

Materials • Taq. Man® Fast Advanced Master Mix • Taq. Man® Gene Expression Assays: • 18 S Hs 99999901_s 1 VIC Primer-limited • GPR 6 Hs 00920605_s 1 FAM • PPIA Hs 99999904_m 1 FAM • YWHAZ Hs 03044281_g 1 FAM • c. DNA: Universal Human Reference (UHR) • Power. Up™ SYBR™ Green Master Mix • Targets (all primers run at final concentration of 300 n. M) • A 2 M ADAM 10 • AOF 1 APOA 1 • ARL 1 C 11 orf 30 • GAPDH PGK 1 • c. DNA: UHR or liver 3

Taq. Man Fast Advanced Master Mix protocol on Bio-Rad CFX 96™ or CFX 384™ (Optional UNG step) https: //tools. thermofisher. com/content/sfs/brochures/cms_087006. pdf Note: Use an anneal temperature appropriate for your assay or primer pair design 4

Taq. Man Fast Advanced Master Mix: Reproducibility From dynamic range data with 3 replicates GAPDH GPR 6 PPIA YWHAZ Cq Std Dev 1. 0 0. 9 0. 8 Cq < 32 0. 7 0. 6 0. 5 0. 4 0. 3 0. 2 0. 1 0. 00 5 5. 00 10. 00 15. 00 20. 00 25. 00 30. 00 35. 00

Taq. Man Fast Advanced Master Mix: Six log dynamic range GAPDH PPIA 6 YWHAZ

Power. Up™ 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 x 40 cycles 60 ºC / 30 sec Melt curve 7

Power. Up™ SYBR™ Green Master Mix: Dynamic Range Excellent efficiency and reproducibility Cq Std Dev 1 0. 9 0. 8 Cq < 32 0. 7 0. 6 0. 5 0. 4 0. 3 0. 2 0. 1 0 0. 00 8 5. 00 10. 00 15. 00 20. 00 25. 00 30. 00 35. 00

Power. Up™ SYBR™ Green Master Mix: PGK 1 R 2 value of 99. 9% shows excellent linearity over a wide input range Single peak showing high target specificity Steep curves show high efficiency 9

Power. Up™ SYBR™ Green Master Mix: ARL 1 R 2 value of 99. 8% shows excellent linearity over a wide input range Single peak showing high target specificity Steep curves show high efficiency 10

Power. Up™ SYBR™ Green Master Mix: Excellent Reproducibility, Six targets A 2 M AOF 1 ARL 1 ADAM 10 APOA 1 C 11 orf 30 11

Power. Up™ SYBR Green Master Mix: Excellent Specificity – Dissociation Data A 2 M ADAM 10 APOA 1 ARL 1 AOF 1 C 11 orf 30 12

Conclusions and Next Steps • Applied Biosystems™ consumables perform well on the Bio-Rad CFX™ (96 & 384 well platforms) • Taq. Man® Gene Expression Assays run with Taq. Man® Fast Advanced Master Mix shows excellent reproducibility, dynamic range and equivalent results in singleplex vs duplex. • Power. Up™ SYBR™ Green Master Mix shows excellent reproducibility and sensitivity. • To learn more about Applied Biosystems q. PCR master mixes or to request a free sample, please visit www. thermofisher. com/qpcrmm • To download the ‘Design and Optimization of SYBR Green Assays’ and/or ‘Protocol for Comparing SYBR Green Master Mixes’ please visit www. thermofisher. com/sybr • Did you know Taq. Man assays are guaranteed to perform? To learn more about Taq. Man assays or purchase an extra small size, please visit www. thermofisher. com/taqman © 2017 Thermo Fisher Scientific Inc. All rights reserved. All trademarks are the property of Thermo Fisher Scientific and its subsidiaries unless otherwise specified. Taq. Man is a registered trademark of Roche Molecular Systems, Inc. , used under permission and license. For research use only. Not for use in diagnostic procedures. 13