QuantiTect Probe PCR Kits
For real-time PCR and two-step RT-PCR using sequence-specific probes
- Highly sensitive detection of low-copy targets
- Accurate quantification over several logs of template
- Use of any sequence-specific probe on any real-time cycler
- Available with or without uracil-N-glycosylase (UNG)
- No need to optimize reaction and cycling conditions
QuantiTect Probe PCR Kits enable sensitive quantification of gDNA and cDNA targets by real-time PCR and two-step RT-PCR using sequence-specific probes. 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 dNTP mix includes dUTP, allowing optional treatment with UNG. For convenience, the master mix can be stored at 2–8°C.
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Cat No./ID:204343 QuantiTect Probe PCR Kit (200) $479.00 For 200 x 50 µl reactions: 3 x 1.7 ml 2x QuantiTect Probe PCR Master Mix, 2 x 2 ml RNase-Free Water |
Cat No./ID:204345 QuantiTect Probe PCR Kit (1000) $1,976.00 For 1000 x 50 µl reactions: 25 ml 2x QuantiTect Probe PCR Master Mix, 20 ml RNase-Free Water |
Cat No./ID:204363 QuantiTect Probe PCR +UNG Kit (200) $581.00 For 200 x 50 µl reactions: 3 x 1.7 ml 2x QuantiTect Probe PCR Master Mix, 100 ul UNG Solution, 2 x 2 ml RNase-Free Water |
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Product Details
The QuantiTect Probe PCR Kit delivers sensitive and reliable results in combination with the QuantiTect Reverse Transcription Kit (see figure "High sensitivity and efficiency, and wide dynamic range"). The unique PCR buffer composition enables QuantiTect Probe PCR Kits to provide sensitive quantification of low-copy DNA targets, as well as accurate quantification over a wide linear range (see figures "High sensitivity and efficiency, and wide dynamic range" and "Wide dynamic range in two-step RT-PCR"). HotStarTaq DNA Polymerase further increases the specificity of the PCR reaction by providing the most stringent hot start compared with other polymerases (see figure "Highly specific amplification").
A major source of PCR contamination is the carryover of PCR products from previous reactions. Pretreatment with UNG prior to starting PCR ensures that any contaminating PCR products do not affect subsequent PCR. The combination of the specially optimized UNG solution and the proven PCR master mix in the QuantiTect Probe PCR +UNG Kit ensures effective elimination of carried-over PCR products together with reliable quantification of target sequences (see figures "Effective UNG digestion" and "Wide dynamic range in real-time PCR").
QuantiTect Probe PCR Kits contain an optimized, ready-to-use master mix for highly specific and sensitive real-time quantification of gDNA and cDNA targets using sequence-specific probes.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), FRET probes, and Molecular Beacons. QuantiTect Probe PCR Kits containa unique PCR buffer that containsa 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 DNA Polymerase provides a stringent hot start, preventingthe formation of nonspecific products.
QuantiTect Probe PCR Master Mix also contains dUTP, enabling pretreatment with uracil-N-glycosylase (UNG) prior to starting PCR, which ensures that any contaminating PCR products do not affect subsequent PCR reactions.
| Component | Features | Benefits |
|---|---|---|
| HotStarTaq DNA Polymerase | 15 minactivation at 95ºC | Set-up of qPCR reactions at room temperature |
| QuantiTect Probe PCR Buffer | Balanced combination of NH4+ and K+ ions | Specific primer annealing ensures reliable PCR results |
| dNTP mix | Includes dUTP, which partially replaces dTTP and enables optional UNG treatment of reactions | Eliminatescontamination from carryover of PCR products by optional UNG treatment |
| ROX dye | For normalization of fluorescent signals on Applied Biosystems and, optionally, Agilent instruments | Precise quantification on cyclers that require ROX dye.Does not interfere with reactions on other real-time cyclers |
QuantiTect Probe PCR Kits overcome the need for optimization of reaction conditions, which can be tedious and time-consuming. Simply add primers, probe, and DNA template to the ready-to-use PCR master mix, and start the reaction (see flowchart "Two-step RT-PCR"). Follow the protocol in the handbook to get fast and reliable results on any real-time cycler. If required, reactions can be pretreated with uracil-N-glycosylase (UNG) to eliminate carryover of PCR products from previous reactions.
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.
QuantiTect Probe PCR Kits can be used for probe-based gene expression analysis of cDNA targets or 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.
Specifications
Features | Specifications |
| Applications | Real-time quantification of DNA, cDNA, or RNA targets |
| Reaction type | PCR and two-step RT-PCR |
| Real-time or endpoint | Real-time |
| Sample/target type | DNA, cDNA, RNA |
| Single or multiplex | Single |
| SYBR Green I or sequence-specific probes | Sequence-specific probes |
| Thermal cycler | Most real-time cyclers (e.g. LC, RG, ABI) |
| With or without ROX | With ROX |
Product Resources
Product citations in scientific articles
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