Description
General description
KAPA SYBR® FAST qPCR Kits contain the first DNA polymerase engineered via directed evolution to be more tolerant of SYBR Green I dye inhibition.
The improved robustness, processivity, and speed of KAPA SYBR FAST qPCR Kits result in consistently high amplification efficiencies enabling more accurate relative quantification for gene expression analysis. KAPA SYBR FAST qPCR Kits, developed to perform optimally in stringent real-time PCR reaction conditions, exhibit dramatic improvements to signal-to-noise ratio (fluorescence), quantification cycle (Cq), linearity, and sensitivity.
Application
KAPA SYBR® FAST for LightCycler®480 has been used for:
Gene expression
Gene knockdown validation
Microarray validation
ChIP analysis
Low copy detection
Absolute quantification of NGS libraries
Quantitative real-time PCR (qPCR)
Features and Benefits
Quantitate changes in gene expression more accurately
High reaction efficiency between 95 - 105% improves accuracy and reproducibility.
Unbiased efficiency across a wide range of GC contents and amplicon lengths.
Detect low copy and difficult targets consistently
Improved processivity results in earlier Cq scores.
Higher fluorescence detection across varying AT- and GC-rich targets.
Novel enzyme is resistant to SYBR® inhibition.
Complete real-time PCR runs in just 40 minutes
55% shorter run times with fast cycling protocol.
Maintain high performance when switching from standard to fast protocols.
Quick Notes:
This kit contains an engineered enzyme optimized for qPCR using SYBR Green I dye chemistry.
The 2X master mix contains a proprietary buffer. Together with the novel enzyme, this improves amplification efficiency of both GC- and AT-rich targets.
20 sec initial denaturation at 95°C is sufficient for enzyme activation. When working with complex templates, an initial denaturation of 3 min is recommended.
For 3-step cycling, use 20 sec for primer annealing and 1 sec for extension/data acquisition at 72°C.
Do not exceed 25 μL reaction volumes.
Preparation Note
Always ensure that components have been fully thawed and thoroughly mixed before use. KAPA SYBR® FAST qPCR Master Mix (2X) may not freeze solidly, even when stored at -20°C.
The SYBR Green I dye contained in KAPA SYBR FAST qPCR Master Mix (2X) and ROX/fluorescein dyes (depending on kit configuration) are light sensitive. Exposure to direct light for an extended period of time will result in loss of fluorescent signal intensity.
KAPA SYBR FAST qPCR Master Mix (2X) is stable through 30 freeze-thaw cycles. Ensure that all reagents are stored protected from light at -20°C when not in use. When protected from light, reagents are stable in the dark at 4°C for at least one week and may be stored at this temperature for short-term use, provided that they do not become contaminated with microbes and/or nucleases.
Analysis Note
All kit components are subjected to stringent functional quality control, are free of detectable contaminating exo and endonuclease activity, and meet strict requirements with respect to DNA contamination.
Other Notes
For Research Use Only. Not for use in diagnostic procedures.
Legal Information
Eppendorf is a registered trademark of Eppendorf SE
LightCycler is a registered trademark of Roche
Rotor-Gene is a registered trademark of Qiagen GmbH
SYBR is a registered trademark of Thermo Fisher Scientific or its subsidiaries
SmartCycler is a registered trademark of Cepheid, Inc.
