QuantiFast Pathogen +IC Kits
For sensitive and reliable detection of viral RNA/DNA and bacterial DNA, including internal control
- Simultaneous detection of pathogen target and internal control
- 5x master mix for higher sensitivity with more sample input
- Correct interpretation of negative results for increased process safety
- Clear detection of weak positive target signals
- Fast, universal protocol on both standard and fast cyclers
The QuantiFast Pathogen +IC Kits are designed for sensitive and rapid real-time PCR or one-step RT-PCR detection of pathogen nucleic acids using sequence-specific probes. To enable high process safety through correct interpretation of negative detection results, each kit contains reagents for multiplex real-time detection of up to 4 user-defined pathogen targets (e.g., virus, bacteria, fungi etc.) plus the Internal Control (IC). Two kit formats are available: The QuantiFast Pathogen RT-PCR +IC Kit for detection of viral RNA, which contains an internal RNA control template, plus the Internal Control primer/probe set, or the QuantiFast Pathogen PCR +IC Kit for detection of viral, bacterial, or fungal DNA, which contains an internal DNA control template, plus the Internal Control primer/probe set. With both kits, ROX is supplied in 2 tubes of different concentrations, enabling use on virtually any real-time instrument. For convenience, the master mix can be stored at 2–8°C.
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Cat No./ID:211352 QuantiFast Pathogen PCR +IC Kit (100) $268.00 For 100 x 25 µl reactions: Master Mix, lyophilized Internal Control Assay, lyophilized Internal Control DNA, ROX Dye Solution, High-ROX Dye Solution, RNase-Free Water, Nucleic Acid Dilution Buffer, Buffer TE |
Cat No./ID:211354 QuantiFast Pathogen PCR +IC Kit (400) $945.00 For 400 x 25 µl reactions: Master Mix, lyophilized Internal Control Assay, lyophilized Internal Control DNA, ROX Dye Solution, High-ROX Dye Solution, RNase-Free Water, Nucleic Acid Dilution Buffer, Buffer TE |
Cat No./ID:211392 Internal Control DNA (High conc.) $76.20 For approximately 200 preps (depending on the elution volume): Lyophilized Internal Control DNA, Nucleic Acid Dilution Buffer |
Cat No./ID:211454 QuantiFast Pathogen RT-PCR +IC Kit (400) $1,129.00 For 400 x 25 µl reactions: Master Mix, RT Mix, lyophilized Internal Control Assay, lyophilized Internal Control RNA, ROX Dye Solution, High-ROX Dye Solution, RNase-Free Water, Nucleic Acid Dilution Buffer, Buffer TE |
Cat No./ID:211492 Internal Control RNA (High conc.) $74.80 For approximately 200 preps (depending on elution volume): Lyophilized Internal Control RNA, Nucleic Acid Dilution Buffer |
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Product Details
QuantiFast Pathogen +IC Kits enable simultaneous detection of viral RNA or DNA targets plus thesupplied Internal Control over a wide linear range without loss of sensitivity when multiplexing (see figures Sensitive detection of Norovirus on the Rotor-Gene Q and High linearity and precision of singleplex and duplex detection). The protocol has been developed to run fast cycling experiments on most cyclers with high reliability (see figures "Sensitive detection of BHV-1 on the Rotor-Gene Q" and "Sensitivedetection ofBHV-1 on theABI 7500"). Combined amplification of pathogen target and the Internal Control using the QuantiFast Pathogen +IC Kits increases process safety of the pathogen detection workflow by ensuring the correct interpretation of negative results (see figure "Correct interpretation of negative results").
QuantiTect Nucleic Acid Dilution Buffer, supplied with the kits, stabilizes RNA and DNA standards during dilution and reaction setup and prevents loss of nucleic acids on plastic surfaces, such as tubes or pipet tips. It enables reliable dilution of standards used to quantify viral nucleic acids, giving a wide linear range, from low to high CT values. The buffer ensures longer storage of standards without degradation (see figure "Reliable dilution and storage of RNA standards").
To enable high process safety through the correct interpretation of negative results, each QuantiFast Pathogen _IC Kit contains reagents for multiplex, real-time detection of a user-defined pathogen target with the Internal Control. Amplifying control and target genes in the same reaction, instead of separate reactions, increases the reliability of gene quantification by minimizing handling errors.
QuantiFast Pathogen +IC Kits provide sensitive and rapid real-time multiplex PCR or one-step RT-PCR detection of pathogen nucleic acids on the first attempt (see flowchart "QIAGEN multiplex kits"). The optimized master mix ensures that PCR products in a multiplex reaction are amplified with the same efficiency and sensitivity as PCR products in a corresponding single-amplification reaction. The specially developed fast PCR buffer contains the novel additive Q-Bond, which significantly reduces denaturation, annealing, and extension times (see figure "Fast primer annealing"). Abalanced combination of K+ and NH4+ ions as well as unique synthetic Factor MP, promote stable and efficient annealing of primers and probes to the nucleic acid template, enabling high PCRefficiency (see figure "Unique PCR buffer"). In addition, theunique formulation of Sensiscript Reverse Transcriptase ensures highly sensitive reverse transcription of viral RNA, while HotStarTaq Plus DNA Polymerase provides a stringent hot start, preventing the formation of nonspecific products.
| Kit component | Features | Benefits | |
|---|---|---|---|
| 5x QuantiFast Pathogen PCR Master Mix | Concentrated master mix | Highly concentrated and optimized for sensitive pathogen detection | Larger volumes of template can be added to the assay for increased sensitivity |
| HotStarTaq Plus DNA Polymerase | 5 min activation at 95ºC | Set up of qPCR reactions at room temperature | |
| QuantiFast PathogenBuffer | Balanced combination of NH4+ and K+ ions | Specific primer annealing ensures reliable PCR results | |
| Synthetic Factor MP | Reliable multiplexing analysis of up to 4 genes in the same tube | ||
| Unique Q-Bond additive | Faster PCR run times, enabling faster results and more reactions per day | ||
| Internal Control Assay | Internal Control template | Internal Control DNA template in the QuantiFast Pathogen PCR +IC Kit | A universalDNA amplification control that can be used with different pathogen assays |
| Internal Control RNA in the QuantiFast Pathogen RT-PCR +IC Kit | A universalRNA amplification control that can be used with different pathogen assays | ||
| Internal Control Assay | Premixed primer/probe set (TaqMan® probe) labeled with MAX (equivalent to HEX, VIC, etc.) | Will not interfere with primers against the pathogen-target | |
| Additional kit components | QuantiFast Pathogen RT Mix* | Contains a unique formulation of Sensiscript Reverse Transcriptase | Optimized for highly sensitive detection of pathogen RNA |
| ROX Dye Solution | Separate tube of passive reference dye for normalization of fluorescent signals on Applied Biosystems 7500 real-time PCR systems. Optional: Can be used on Stratagene instruments from Agilent | Precise quantification on cyclers that require ROX dye.Does not interfere with PCR on any real-time cycler | |
| High-ROX Dye Solution | Separate tube of passive reference dye for normalization of fluorescent signals on Applied Biosystems 7900 and StepOne real-time PCR systems | ||
| QuantiTect Nucleic Acid Dilution Buffer | Proprietary buffer formulation for dilution and storage of nucleic acid standards | Stabilizes RNA and DNA standards during dilution and reaction setup and prevents loss of nucleic acids on plastic surfaces, such as tubes or pipet tips |
QuantiFast Pathogen +IC Kits provide a simple procedure for the detection of a user-defined pathogen and the Internal Control. They contain a ready-to-use master mix for the real-time detection of viral RNA (1-step RT-PCR) or viral, bacterial, and fungalDNA (PCR). There is no need for optimization of reaction and cycling conditions. Simply mix the supplied master mix with the pathogen assay (primers and probe) and the supplied Internal Control Assay and the Internal Control DNA or RNA. Alternatively, if the InternalControl has been added to the sample purification procedure, add RNase-free water instead of Internal Control DNA or RNA to the reaction mix. Then add your sample DNA or RNA and start the reaction on any cycler. The kit handbook contains optimized protocols for use with TaqMan® probes on a wide range of cyclers. It also contains recommendations for selection of dye combinations.
Each QuantiFast Pathogen +IC Kit includes the Internal Control Assay and Internal Control DNA or RNA for use as an amplification control via direct addition to the reaction mix. Alternatively, the IC can be added to the purification procedure to control both the purification process and amplification. For addition of the Internal Control to the purification procedure, highly concentrated Internal Control DNA or RNA (High conc.) can be ordered separately (see figure "QIAGEN Internal Control").
QuantiFast Pathogen +IC Kits provide sensitive real-time PCR or one-step RT-PCR using sequence-specific probes for detection of pathogen DNA and/or RNA including an internal control. The kits can be used on a wide range of real-time cyclers, including cyclers from QIAGEN, Applied Biosystems, Bio-Rad, Roche (except for capillary cyclers), and Agilent.
Specifications
Features | Specifications |
| Applications | Pathogen Detection: Real-time PCR of viral, bacterial or fungal DNA (QuantiFast Pathogen PCR +IC Kit) or one-step RT-PCR for detection of viral RNA (QuantiFast Pathogen RT-PCR +IC Kit) |
| Reaction type | Real-time PCR or one-step RT-PCR including of an internal control (IC) |
| Real-time or endpoint | Real-time |
| Sample/target type | QuantiFast Pathogen PCR +IC Kit: viral, bacterial or fungal DNA; QuantiFast Pathogen RT-PCR +IC Kit: viral RNA |
| Single or multiplex | Duplex |
| SYBR Green I or sequence-specific probes | Sequence-specific probes |
| Thermal cycler | For most standard and fast real-time cylcers compatible with duplex PCR/RT-PCR, e.g. Rotor-Gene Q or cyclers from Agilent, Applied Biosystems, BioRad, Roche |
| With or without ROX | Master Mix is provided without ROX dye, but 2 separate ROX solutions are included: High-ROX Dye Solution for use with ABI cyclers except ABI 7500, ROX Dye Solution (low ROX conc.) for use with ABI 7500 and other suppliers |
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