KAPA SYBR® FAST qPCR Kits

Evolved for the Highest Performance in Real Time PCR

KAPA SYBR® FAST qPCR Master Mix is designed for the highest performance in 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 KAPA SYBR® DNA Polymerase has been engineered to perform optimally in stringent real-time qPCR reaction conditions, exhibiting dramatic improvements to signal-to-noise ratio (fluorescence), cycle threshold (CT), linearity, and sensitivity. The KAPA SYBR® DNA Polymerase and proprietary buffer system enhances the amplification efficiency of difficult templates, including both GC-rich and AT-rich templates.

KAPA SYBR® FAST qPCR Kits are ideally suited for:

• Gene expression analysis
• Low copy gene detection
• Microarray validation
• Gene knockdown validation
• High-throughput qPCR

Superior signals, Ct values, and reaction efficiencies

The high performance of the KAPA SYBR® FAST qPCR kit can been seen when compared to other leading Fast qPCR Master Mixes. KAPA SYBR® FAST consistently produces higher signal, earlier Cts, and 100% reaction efficiencies.

Human B-actin (290 bp) was amplified from log-fold dilutions of human genomic DNA (20 ng to 2 pg) using KAPA SYBR® FAST qPCR Master Mix (2x) optimized for Roche LightCycler® 480 or Roche LightCycler® 480 SYBR Green I Master kit (Roche). Comparisons were performed according to the competitor protocol A: Roche protocol: 95°C, 5 min followed by 40 cycles of 95°C, 10 sec; 60°C, 15 sec; 72°C, 6 sec; or B: KAPA SYBR® FAST Protocol: 95°C 3 min followed by 40 cycles of 95°C, 10 sec; 60°C, 15 sec; 72°C, 1 sec.
 

High speed and high performance

High speed does not compromise performance with the KAPA SYBR® FAST qPCR kits. As can been seen from the amplification plots and standard curves there is no loss in performance between fast and conventional cycling conditions even with complex targets.

 
Human coagulation factor V (F5) was amplified from log-fold dilutions of human genomic DNA (20 ng-2 pg) using KAPA SYBR® FAST qPCR Master Mix, with fast cycling protocol, or with conventional cycling conditions. There is no loss in performance between fast and conventional cycling conditions even with complex targets. Although the proprietary antibody hot start is activated after 20 seconds at 95°C, 3 min initial activation at 95°C is recommended for optimal denaturation of complex targets.

Highest performance on Roche LightCycler® 480
KAPA SYBR® FAST qPCR Master Mix (2x) is specifically formulated for the highest performance on the Roche LightCycler® 480 instrument. A dramatic loss in performance of the Roche LightCycler® 480 SYBR® Green I Master kit was observed as the extension time at 72°C was reduced from 6 sec to 1 sec. There was no loss in performance of the KAPA SYBR® FAST qPCR Master Mix (2x) Kit optimized for the Roche LightCycler® 480.

 Human B-actin (290 bp) was amplified from log-fold dilutions of human genomic DNA (20 ng to 2 pg) using KAPA SYBR® FAST qPCR Master Mix (2x) Roche LightCycler® 480 or Roche LightCycler® 480 SYBR Green I Master kit (Roche). Comparisons were performed according to the competitor protocol A: Roche protocol: 95°C, 5 min followed by 40 cycles of 95°C, 10 sec; 60°C, 15 sec; 72°C, 6 sec; or B: KAPA SYBR® FAST Protocol: 95°C 3 min followed by 40 cycles of 95°C, 10 sec; 60°C, 15 sec; 72°C, 1 sec.

Highest fluorescence and earliest Ct

The KAPA SYBR® FAST qPCR kit displays the highest fluorescence and earliest Ct values from a range of AT and GC-rich amplicons.

Human B-actin, human coagulation factor V (F5), and HPRT1 targets were amplified from 20ng (5900 copies) of human genomic DNA on an ABI Prism® 7900 HT sequence detection system. Values represent an average of triplicates.
 

KAPA SYBR® FAST qPCR Kits are intended for use on all real-time instruments and protocols.

KAPA SYBR® FAST qPCR Master Mix (2x) is a ready-to-use cocktail containing all components except primers and template, for the amplification and detection of DNA in qPCR.

The KAPA SYBR® FAST qPCR Kit is supplied as a 2x Master Mix with integrated antibody-mediated hot start, SYBR® Green I fluorescent dye, MgCl2, dNTPs, and stabilizers. ROX reference dye is included in the ABI Prism® Master Mix (2X), and supplied in separate tubes in the Universal Master Mix (2X). Fluorescein reference dye is included in the Bio-Rad iCycler Master Mix (2x). The KAPA SYBR® FAST qPCR Master Mix (2X) optimized for the Roche LightCycler® 480 does not contain reference dye.

Quick OrderSYBR® is a registered trademark of Molecular Probes, Inc, Oregon. LightCycler® is a registered trademark of Roche. PRISM® is a registered trademark of Appelera. iCycler® is a registered trademark of Bio-Rad.

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