QuantiTect Probe RT-PCR Kit
For one-step qRT-PCR using sequence-specific probes for gene expression analysis
- 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
- No need to optimize reaction and cycling conditions
QuantiTect Probe RT-PCR Kits enable sensitive quantification of RNA targets by real-time one-step 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 qRT-PCR 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:204443 QuantiTect Probe RT-PCR Kit (200) $672.00 For 200 x 50 µl reactions: 3 x 1.7 ml 2x QuantiTect Probe RT-PCR Master Mix, 100 µl QuantiTect RT Mix, 2 x 2 ml RNase-Free Water |
Cat No./ID:204445 QuantiTect Probe RT-PCR Kit (1000) $2,763.00 For 1000 x 50 µl reactions: 25 ml 2x QuantiTect Probe RT-PCR Master Mix, 0.5 ml QuantiTect RT Mix, 20 ml RNase-Free Water |
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Product Details
The QuantiTect Probe RT-PCR Kit has a unique RT-PCR buffer that promotes highly specific annealing of primers and probes to the PCR template. It contains HotStarTaq DNA Polymerase, which provides the most stringent hot start compared with other polymerases (see figure "Highly specific amplification").
HotStarTaq DNA Polymerase and the unique composition of the RT-PCR buffer enable the QuantiTect Probe RT-PCR Kit to provide sensitive quantification of low-copy RNA targets, as well as accurate quantification over a wide linear range (see figure "High sensitivity and efficiency, and wide dynamic range"). Performing reverse transcription and PCR sequentially in the same tube does not impair sensitivity, as demonstrated by CT values that are comparable to those achieved in real-time two-step RT-PCR (see figures "One-step RT-PCR with comparable performance to two-step RT-PCR — A" and "One-step RT-PCR with comparable performance to two-step RT-PCR — B", and table).
| Amount of cDNA/RNA | Mean CT value | Mean CT value |
|---|---|---|
| (two-step RT-PCR) | (one-step RT-PCR) | |
| 100 ng | 24.66 | 24.66 |
| 10 ng | 28.15 | 27.84 |
| 1 ng | 31.42 | 31.49 |
| 0.1 ng | 34.80 | 34.56 |
| 0.01 ng | 37.78 | 37.66 |
QuantiTect Probe RT-PCR Kits contain an optimized, ready-to-use master mix for highly specific and sensitive real-time quantification of RNA targets using sequence-specific probes.The kits are designed for use with all types of sequence-specific probes, including hydrolysis probes (e.g., TaqMan® and other dual-labeled probes), FRET probes, and Molecular Beacons. QuantiTect Probe RT-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, anoptimized mix of reverse transcriptasesenables cDNA synthesis from a wide range of RNA template amounts, whileHotStarTaq DNA Polymerase provides a stringent hot start, preventingthe formation of nonspecific products.
QuantiTect Probe RT-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 RT-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 |
| Omniscript and Sensiscript Reverse Transcriptases | Special blend of enzymes with high affinity for RNA | RNA can be transcribed, even through complex secondary structures |
The QuantiTect Probe RT-PCR Kit overcomes the need for optimization of reaction conditions, which can be tedious and time-consuming. Simply add primers, probe, and RNA template to the ready-to-use RT-PCR master mix, and start the reaction (see flowchart "One-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.
QuantiTect Probe RT-PCR Kits can be used for 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
Features | Specifications |
| Applications | Real-time quantification of RNA targets |
| Reaction type | One-step RT-PCR |
| Real-time or endpoint | Real-time |
| Sample/target type | RNA targets |
| Single or multiplex | Single |
| SYBR Green I or sequence-specific probes | Sequence-specific probes |
| Thermal cycler | All 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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