QuantiFast Probe PCR Kits

For fast, real-time PCR and two-step qRT-PCR using sequence-specific probes

Products

QuantiFast Probe PCR Kits are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.
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QuantiFast Probe PCR Kit (2000)

Cat. No. / ID. / ID.   204256

IMPORTANT NOTE: As announced earlier, the production of the QuantiFast kits has been discontinued since mid-2021. Hence, these products will be available only until stocks last.   For 2000 x 25 µl reactions: 25 ml 2x QuantiFast Probe PCR Master Mix (contains ROX dye), 20 ml RNase-Free Water
50.711,00 MXN
This kit is being phased out. We recommend switching to the QuantiNova successor product. For more information and FAQs on this transition, visit: www.qiagen.com/PCRresource.

Features

  • Sensitive detection of even low copy targets
  • Accurate detection of a wide range of template amounts
  • Faster results with a time savings of up to 60%
  • Optimized, ready-to-use master mix for reliable results without optimization
  • One cycling protocol for all standard and fast cyclers

Product Details

QuantiFast Probe PCR Kits deliver fast and sensitive quantification of gDNA and cDNA targets by real-time PCR and two-step RT-PCR using sequence-specific probes. Q-bond technology and an optimized master mix enable shorter real-time 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. Two kit formats are available: the QuantiFast Probe PCR Kit for cyclers that require ROX dye for fluorescence normalization, and the QuantiFast Probe PCR +ROX Vial Kit for all other cyclers. For convenience, the master mix can be stored at 2–8°C.

IMPORTANT NOTE: As announced earlier, the production of the QuantiFast kits has been discontinued since mid-2021. Hence, these products will be 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.

Performance

QuantiFast Probe PCR Kits deliver highly sensitive results, outperforming other real-time PCR kits (see table "Sensitive two-step RT-PCR on the iCycler iQ" and figure and  Sensitive two-step RT-PCR). 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 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.

Sensitive two-step RT-PCR on the iCycler iQ
  Mean CT value  
Human leukocyte cDNA (ng) QuantiFast Probe PCR +ROX Vial Kit Kit from Supplier BV
100 29.13 31.13
10 32.50 34.43
1 36.15 40.00
No template control 40.00 40.00

 

QuantiFast Probe PCR Kits allow accurate quantification over a wide dynamic range (see figure " Wide dynamic range and high sensitivity"). Template dilutions of up to 7 logs can be reliably detected (see table "Detection over 7 logs of template in two-step RT-PCR on the LightCycler 2.0 " and figure " Detection over 7 logs of template").

Detection over 7 logs of template in two-step RT-PCR on the LightCycler 2.0
  Mean CT value  
Human leukocyte cDNA QuantiFast Probe PCR +ROX Vial Kit Kit from Supplier R
100 ng 11.99 14.56
10 ng 15.54 17.99
1 ng 19.22 22.16
100 pg 22.63 25.14
10 pg 26.43 30.15
1 pg 30.02 34.43
100 fg 33.42 37.10
No template control 40 40.00
PCR efficiency 90% 80.1%
See figures

Principle

QuantiFast Probe 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. A specially developed, fast PCR buffer contains the novel additive Q-Bond, which significantly reduces denaturation, annealing, and extension times (see figure " Fast primer annealing"). The buffer also contains a balanced combination of K+ and NH4+ ions, which promote specific primer annealing, enabling high PCR specificity and sensitivity (see figure " Specific primer annealing"). In addition, HotStarTaq Plus DNA Polymerase provides a stringent hot start, preventing the formation of nonspecific products.

Components of 2x QuantiFast Probe PCR Kit*
Component Features and benefits Benefits
HotStarTaq Plus DNA Polymerase 3 min activation at 95ºC Set up of qPCR reactions at room temperature
QuantiFast Probe PCR Buffer Balanced combination of NH4+ and K+ ions Specific primer annealing ensures reliable PCR results
Unique Q-Bond additive Faster PCR run times, enabling faster results and more reactions per day
ROX dye Normalizes fluorescent signals on Applied Biosystems and, optionally, Agilent instruments Precise quantification on cyclers that require ROX dye. Does not interfere with PCR on any real-time cycler
See figures

Procedure

QuantiFast Probe PCR Kits contain ready-to-use master mixes that eliminate the need for optimization of reaction and cycling conditions. Simply add template DNA, 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.

Choosing the right QuantiFast Probe PCR Kit
ROX dye Kit Compatible cyclers
Supplied in master mix QuantiFast Probe PCR Kit All cyclers from Applied Biosystems except Applied Biosystems 7500
Supplied in a separate tube QuantiFast Probe PCR +ROX Vial Kit Applied Biosystems 7500, and cyclers from Agilent, Bio-Rad, Cepheid, QIAGEN, Eppendorf, Roche, and other suppliers

For optimal results in real-time two-step RT-PCR, we recommend synthesizing cDNA using the QuantiTect Reverse Transcription Kit. The kit provides fast cDNA synthesis in just 20 minutes with integrated removal of genomic DNA contamination. 

QuantiFast Probe Assays are predesigned, genomewide assays that use hydrolysis, probe-based detection. They are delivered with the QuantiFast Probe PCR Kit for guaranteed results in singleplex, two-step qRT-PCR.

Applications

QuantiFast Probe PCR Kits can be used for probe-based gene expression analysis of cDNA targets and quantitative gDNA analysis 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 PCR Kit, which has been specially developed for fast cycling on these instruments.

Supporting data and figures

Specifications

FeaturesSpecifications
applicationsProbe-based, real-time PCR, two-step RT-PCR
sampletargettypecDNA, DNA
realtimeorendpointReal-time
reactiontypeReal-time and two-step RT-PCR
singleormultiplexSingle
withorwithoutroxAvailable with ROX in master mix and with ROX as separate vial
sybrgreeniorsequencespecificprobesSequence-specific probes
thermalcyclerAll real-time cyclers (e.g. Roche LightCycler®, Corbett Rotor-Gene, Applied Biosystems)

Resources

Safety Data Sheets (1)
Download Safety Data Sheets for QIAGEN product components.
Quick-Start Protocols (2)
Kit Handbooks (1)
For fast, quantitative, real-time PCR and two-step RT-PCR using sequence-specific probes
Certificates of Analysis (1)