QuantiFast Probe RT-PCR Kits
For fast, one-step qRT-PCR using sequence-specific probes for gene expression analysis
QuantiFast Probe RT-PCR Kits deliver fast and sensitive real-time one-step RT-PCR quantification of RNA targets using probe-based detection. Q-bond technology and an optimized master mix enable shorter real-time RT-PCR run times, not only on fast cyclers with short ramping times, but also on standard cyclers. The combination of a hot start and a unique PCR buffer system in the ready-to-use master mix ensures highly sensitive qPCR on any real-time cycler without the need for optimization. The kits are also supplied with an optimized RT mix for efficient cDNA synthesis in only 10 minutes. Two kit formats are available: the QuantiFast Probe RT-PCR Kit for cyclers that require ROX dye for fluorescence normalization, and the QuantiFast Probe RT-PCR +ROX Vial Kit for all other cyclers. For convenience, the master mix can be stored at 2–8°C.
IMPORTANT NOTE: This product is phasing-out until June 30, 2021, and afterward, available only until stocks last. Visit the product page of the successor kit to view improved features or to request a trial kit.
For more information and FAQs on this transition, visit: www.qiagen.com/PCRresource. Buy Products
QuantiFast Probe RT-PCR Kits are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.
Product Details
Performance
QuantiFast Probe RT-PCR Kits deliver highly sensitive results, outperforming other real-time RT-PCR kits (see figure "Sensitive One-step RT-PCR"). RT-PCR run times are reduced by up to 60% (see figure "Significantly reduced PCR times"), allowing you to achieve fast PCR results without compromising on RT-PCR performance (see figure "Faster results without compromising sensitivity"). You can also greatly increase your sample throughput or efficiently share a cycler with other users. With QuantiFast Probe RT-PCR Kits, you follow the same procedure whether you work with a standard or fast cycler. QuantiFast Probe RT-PCR Kits allow accurate quantification over a wide dynamic range (see figure "Comparable dynamic range"). Template dilutions of up to 6 logs can be reliably detected (see figure "Wide dynamic range and high sensitivity").
Principle
QuantiFast Probe RT-PCR Kits deliver highly sensitive and rapid results over a wide dynamic range on both standard and fast cyclers. The kits are designed for use with all types of sequence-specific probes, including hydrolysis probe detection (e.g., TaqMan and other dual-labeled probes) and FRET probes. The optimized QuantiFast RT Mix enables cDNA synthesis in just 10 minutes and a specially developed fast RT-PCR buffer contains the novel additive Q-Bond, which significantly reduces denaturation, annealing, and extension times (see figure "Fast primer annealing"). The RT-PCR buffer also contains a balanced combination of K+ and NH4+ ions, which promote specific primer annealing and enable high RT-PCR specificity and sensitivity (see figure "Specific primer annealing"). In addition, HotStarTaq Plus DNA Polymerase requires only 5 minutes at 95°C for activation and provides a stringent hot start, preventing the formation of nonspecific products.
Procedure
QuantiFast Probe RT-PCR Kits contain ready-to-use master mixes that eliminate the need for optimization of reaction and cycling conditions. Simply add template RNA, primers, and probe to the master mix and follow the protocol in the handbook to get fast and reliable results on any real-time cycler. Kits are available with or without ROX passive reference dye in the master mix, enabling use on virtually any real-time cycler (see table). Due to the optimized ROX concentrations, detection of even low copy numbers is achieved through automatic data analysis.
Applications
QuantiFast Probe RT-PCR Kits can be used for probe-based gene expression analysis of RNA targets on any real-time cycler. This includes instruments from Applied Biosystems, Bio-Rad, Cepheid, Eppendorf, Roche, and Agilent. For the Rotor-Gene Q and other Rotor-Gene cyclers, we recommend using the Rotor-Gene Probe RT-PCR Kit, which has been specially developed for fast cycling on these instruments. Specifications
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