www.kapabiosystems.com

Introducing KAPA™ SYBR® FAST qPCR Kits

KAPA SYBR® FAST qPCR Kits contain a novel DNA polymerase evolved specifically for real-time PCR.

The result is industry-leading performance:

Highest reaction efficiency. Highest signal intensity. Highest sensitivity and linearity. High speed reduces total cycling time.

next generation thinking

in enzyme technology

Containing the first DNA polymerase evolved for real-time PCR.

Amplification Plot

4.000 E+1

9.000 E+1

1.400 E+2

1.900 E+2

2.400 E+2

2.900 E+2

3.400 E+2

∆R

0 5 10 15 20 25 30 35 40Cycle

Kit can be used for this cycler but may require the addition of ROX reference dye

Product Description

KAPA SYBR® FAST qPCR Master Mix is designed for high performance real-time PCR. The kit contains a novel DNA polymerase engineered through a process of molecular evolution. The result is a unique enzyme, specifically evolved for qPCR using SYBR® Green I dye chemistry.

The reaction efficiency of qPCR is critical for accurate and reliable gene quantification. Wild type Taq DNA polymerase is often inhibited by real-time PCR reaction conditions, leading to significant loss in specificity and sensitivity of the assay. The KAPA SYBR® DNA Polymerase has been engineered to perform optimally in stringent qPCR reaction conditions, exhibiting dramatic improvements in signal-to-noise ratio (fluorescence), cycle threshold (CT), linearity, and sensitivity. The novel DNA polymerase and proprietary buffer system enhances the amplification efficiency of difficult templates, including both GC and AT-rich templates.

High speed is achieved through the increased processivity of the engineered KAPA SYBR® DNA Polymerase and the short initial denaturation time required for the activation of the proprietary antibody-mediated hot start.

>> KAPA™SYBR® FAST qPCR Kit Containing the first DNA polymerase evolved for real-time PCR.

Engineered KAPA SYBR® DNA Polymerase and optimized buffer system provides:

Highest reaction efficiency resulting in the earliest cycle thresholds (CT) and extreme sensitivity.

Highest fluorescence resulting in improved signal-to-noise ratio.

Broad dynamic range and linearity.

High speed reduces total cycling time.

Human coagulation factor V (F5) was amplified from log-fold dilutions of human genomic DNA (20 ng - 2 pg) using KAPA SYBR® FAST Master Mix (2x), with standard cycling protocol (green) and fast cycling pro-tocol (blue). Standard cycling conditions: 5 min initial denaturation at 95ºC followed by 40 cycles of 15 sec denaturation at 95ºC and 60 sec combined annealing/extension at 60ºC. Fast cycling conditions: 3 min initial denaturation at 95ºC followed by 40 cycles of 1 sec denaturation at 95ºC and 20 sec combined annealing/extension at 60ºC. Total run time was 1.5 hours for standard cycling and 40 mins for fast cycling.

High speed and high performance

161820222426

101 102 103 104

Ct

Concentration

Standard curveSlope -3.305R2 0.995PCR E�. 101%

1816

2022242628

101100 102 103 104

Ct

Concentration

Standard curveSlope -3.377R2 0.990PCR E�. 98%

Threshold

10-1.0

10-1.2

10-1.4

10-1.6

10-1.8

10-0.8

1.0-0.6

10-0.4

10-0.2

100.0

Norm

. Flu

oro.

5 10 15 20 25 30 35 40Cycle

Amplification Plot

High speed does not compromise performance with the KAPA SYBR® FAST qPCR Kits. As can be seen from the amplification plots and standard curves, high reaction efficiencies are achieved with both fast and standard cycling conditions, even with complex genomic targets. Total run time for the fast cycling protocol was 40 minutes.

Product Applications

KAPA SYBR® FAST qPCR Kits are ideally suited for: Gene expression analysis Microarray validation Low copy gene detection Gene knockdown validation

The KAPA SYBR® FAST qPCR Kit is available as a ready-to-use 2x Master Mix containing all components, except primers and template, for the amplification and detection of DNA in real-time quantitative PCR (qPCR). All KAPA SYBR® qPCR FAST Kits are supplied with antibody-mediated hot start for high specificity and maximum speed.

The KAPA SYBR® FAST qPCR Master Mix(2x) LightCycler™ 480 Kit is formulated for high performance on the Roche LightCycler™ 480 instrument. The KAPA SYBR® FAST qPCR Kit outperforms the competitor using both the recommended cycling protocol and a faster cycling protocol indicating robust, high speed performance.

High performance on Roche LightCycler™ 480

Human B-actin (290 bp) was amplified from 5 log-fold dilutions of human genomic DNA (20 ng - 2 pg) using KAPA SYBR® FAST qPCR Master Mix (2x) LightCycler™ 480 Kit (green) and competitor R (red). Compari-sons were performed according to the competitor R protocol (left panel: 5 min initial denaturation at 95ºC followed by 40 cycles of 10 sec denaturation at 95ºC, 15 sec annealing at 60ºC, 6 sec extension at 72ºC) and KAPA SYBR® FAST protocol (right panel: 3 min initial denaturation at 95ºC followed by 40 cycles of 10 sec denaturation, 15 sec annealing at 60ºC, 1 sec extension at 72ºC).

Superior signal and reaction efficiency

The KAPA SYBR® FAST qPCR Kit outperfoms when compared to other leading fast qPCR kits. The KAPA SYBR® FAST qPCR Kit consistently produces higher signals, earlier Cts, and 100% reaction efficiencies.

Human B-actin (290 bp) was amplified from 5 log-fold dilutions of human genomic DNA (20 ng to 2 pg) using KAPA SYBR® FAST qPCR Master Mix (2x) or fast qPCR competitor kits. KAPA SYBR® FAST qPCR Master Mix (green) vs. Competitor A (orange), competitor Q (purple), or competitor I (blue). Competitor I also amplifies the no template control (NTC). Reactions were performed according to each competitors’ suggested protocol on an ABI Prism® 7900 HT sequence detection system.

Fluor

esce

nce (

483-

533)

Amplification Curves

Cycles21 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40

26.584

24.084

21.584

19.084

16.584

14.084

11.584

9.084

6.584

4.084

1.584

-0.916

KAPA SYBR® FAST Competitor R Slope -3.481 -6.00R2 0.997 0.999PCR E�. 94% 46%

Slope -3.481 -6.00R2 0.997 0.999PCR E�. 94% 46%

Ct

Quantity

4038

34

30

26

22

36

32

28

24

201.50E+00 1.50E+01 1.50E+02 1.50E+03 1.50E+04

Amplification Plot

4.000 E+1

9.000 E+1

1.400 E+2

1.900 E+2

2.400 E+2

2.900 E+2

3.400 E+2

∆R

0 5 10 15 20 25 30 35 40Cycle

Slope -3.331 -3.180R2 0.997 0.994PCR E�. 100% 106%

KAPA SYBR® FAST Competitor A

Ct

Quantity

37

3335

2932

2325

2021

1719

1.50E+00 1.50E+01 1.50E+02 1.50E+03 1.50E+04

Amplification Plot

4.000 E+1

9.000 E+1

1.400 E+2

1.900 E+2

2.400 E+2

2.900 E+2

3.400 E+2

∆R

0 5 10 15 20 25 30 35 40Cycle

Slope -3.331 -3.123R2 0.997 0.983PCR E�. 100% 109%

KAPA SYBR® FAST Competitor Q

Ct

Quantity

37

3335

2932

2325

2021

1719

1.50E+00 1.50E+01 1.50E+02 1.50E+03 1.50E+04

Amplification Plot

5.000 E+1

1.000 E+2

1.500 E+2

2.000 E+2

2.500 E+2

3.000 E+2

3.500 E+2

4.000 E+2

∆R

0 5 10 15 20 25 30 35 40Cycle

NTC

Slope -3.331 -2.881R2 0.997 0.994PCR E�. 100% 122%

KAPA SYBR® FAST Competitor I

Ct

Quantity

37

3335

2932

2325

2021

1719

1.50E+00 1.50E+01 1.50E+02 1.50E+03 1.50E+04

Fluor

esce

nce (

483-

533)

Amplification Curves

Cycles21 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40

24.933

22.433

19.933

17.433

14.933

12.433

9.933

7.433

4.933

2.433

-0.067

KAPA SYBR® FAST Competitor R Slope -3.381 -3.022R2 0.998 0.998PCR E�. 98% 77%

Slope -3.381 -3.022R2 0.998 0.998PCR E�. 98% 77%

Ct

Quantity

4038

34

30

26

22

36

32

28

24

201.50E+00 1.50E+01 1.50E+02 1.50E+03 1.50E+04

www.kapabiosystems.com

Boston, Massachusetts, United States600 West Cummings Park, Suite 5350

Woburn, MA, 01801 U.S.A.Tel: +1 781 497 2933 Fax: +1 781 497 2934

Email: info@kapabiosystems.com

Cape Town, South Africa2nd Floor, Old Warehouse Building, Black River Park,Fir Road, Observatory, 7925 Cape Town, South Africa

Tel: +27 21 448 8200 Fax: +27 21 448 6503Email: info@kapabiosystems.com

Highest fluorescence and earliest CT values

The KAPA SYBR® FAST qPCR Kit exhibits industry-leading performance in both reaction efficiency and signal intensity across a broad range of targets as compared with fast qPCR competitor kits.

Instrument Compatibility

KAPA SYBR® FAST qPCR Kits are supplied as a ready-to-use Master Mix (2x) containing all components, except primers and template, for the amplification and detection of DNA in real-time quantitative PCR (qPCR). The Master Mix (2x) includes hot start KAPA SYBR® DNA polymerase, SYBR® Green I fluorescent dye, MgCl2, dNTPs, reference dye (when applicable), and stabilizers.

For more information on KAPA SYBR® FAST qPCR Kits please visit www.kapabiosystems.comFor technical support please contact: support@kapabiosystems.com

SYBR® is a registered trademark of Molecular Probes, Inc, OregonPRISM®, StepOne™, StepOnePlus™, and GeneAmp® are registered trademarks of Applera Corporation.iCycler®, MyiQ™, iQ™5, Mx3000P®, Mx3005™, Mx4000®, Roto-Gene™, DNA Engine Opticon™, Chromo 4™, LightCycler®, MasterCycler®,and Smart Cycler® are trademarks or registered trademarks of their respective companies.

24

23

22

21

20

19

18

17

16

15

Ct

Comparison of Ct Values

B-actin F5 HPRT1 Amplicon

KAPA SYBR® FAST

Competitor I

Competitor A

Competitor R

Competitor Q

350

300

250

200

150

100

50

0 B-actin F5 HPRT1 Amplicon

Base

line s

ubtra

cted

�uo

resc

ence

Fluorescence Intensity

KAPA SYBR® FAST

Competitor I

Competitor A

Competitor R

Competitor Q

Human B-actin, F5, and HPRT1 targets were amplified from 20 ng (5900 copies) of human genomic DNA on an ABI Prism® 7900 HT sequence detection system. Values represent averages of triplicates. Reactions were performed according to each competitors’ suggested protocol.

KAPA SYBR® FAST qPCR Master Mix (2x) Universal Kit: Includes separate tubes of ROX passive reference dye (high and low)

KAPA SYBR® FAST qPCR Master Mix (2x) ABI Prism® Kit: ROX passive reference dye included in 2x Master Mix

KAPA SYBR® FAST qPCR Master Mix (2x) Bio-Rad iCycler™ Kit: Fluorescein reference dye included in 2x Master Mix

KAPA SYBR® FAST qPCR Master Mix (2x) LightCycler® 480 Kit: Contains no reference dye

KK4600

KK4601

KK4602

KK4603

KK4604

KK4605

KK4606

KK4607

KK4608

KK4609

KK4610

KK4611

1 x 1 ml (100, 20µl reactions)

1 x 5 ml (500, 20µl reactions)

2 x 5 ml (1000, 20µl reactions)

1 x 1 ml (100, 20µl reactions)

1 x 5 ml (500, 20µl reactions)

2 x 5 ml (1000, 20µl reactions)

1 x 1 ml (100, 20µl reactions)

1 x 5 ml (500, 20µl reactions)

2 x 5 ml (1000, 20µl reactions)

1 x 1 ml (100, 20µl reactions)

1 x 5 ml (500, 20µl reactions)

2 x 5 ml (1000, 20µl reactions)

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• •••••••••

•••••••

•••

º º º º º º

• Prefered kit for this cycler º Kit can be used for this cycler but may require the addition of ROX reference dye

•