Description
General description
KAPA PROBE FAST qPCR Kits provide fast and reproducible results for all probe-based qPCR applications. Kits contain a ready-to-use master mix for highly sensitive and accurate real-time PCR using sequence-specific fluorogenic probe chemistries, including hydrolysis probes (e.g. TaqMan®), FRET probes, and displacement probes (e.g. molecular beacons).
Application
KAPA PROBE FAST may be used for:
Real-time quantitative methylation-specific PCR (qMSP)
Genotyping
Gene expression analysis
Multiplex qPCR and qRT-PCR
SNP genotyping/allelic discrimination
Microarray validation
Features and Benefits
Accurately discriminate between alleles:
Exceptional differentiation of heterozygous and homozygous alleles
Discrete clusters in SNP genotyping assays
Achieve excellent reproducibility and efficiency:
Fast, high performance five-colored multiplex PCR
Similar abundance levels achieved with low copy number
Broad dynamic range of up to 10 orders of magnitude
Increase flexibility:
Compatible with both standard and fast cycling protocolsRetain accurate quantification
Quick Notes:
These kits are compatible with all fluorogenic probebased technologies, including hybridization probes (e.g., FRET), hydrolysis probes (e.g., TaqMan®) and displacement probes (e.g., molecular beacons).
Initial denaturation for 20 sec at 95ºC is sufficient for enzyme reactivation; however, optimal denaturation of complex targets may require up to 3 min denaturation.
For 2-step cycling, use 30 sec combined annealing/ extension/data acquisition.
For 3-step cycling, use 20 sec for primer annealing and 1 sec for extension/data acquisition.
Do not exceed 25 μL volumes.
Preparation Note
ROX/fluorescein reference 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. Avoid repeated freezing and thawing. Always ensure that the product has been fully thawed and mixed before use.
KAPA PROBE 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
KAPA PROBE FAST qPCR Master Mix (2X) is subjected to stringent quality control tests, is free of contaminating exoand endonuclease activity, and meets strict requirements with respect to DNA contamination levels.
Other Notes
For Research Use Only. Not for use in diagnostic procedures.
Legal Information
Applied Biosystems is a registered trademark of Applera Corporation or its subsidiaries in the US and/or certain other countries
QuantStudio is a trademark of IROA Technologies LLC
StepOne is a trademark of Applera Corporation or its subsidiaries in the US and/or certain other countries
StepOnePlus is a trademark of Applera Corporation or its subsidiaries in the US and/or certain other countries
TaqMan is a registered trademark of Roche Molecular Systems, Inc.
