QuantiFast SYBR® Green RT-PCR Kit
For fast, one-step qRT-PCR using SYBR Green I for gene expression analysis
- Faster results with time savings of up to 60%
- Sensitive detection of even low copy numbers
- Accurate detection of a wide range of template amounts
- Optimized, ready-to-use master mix for fast cycling
- Universal protocol for all standard and fast cyclers
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Cat No./ID:204154 QuantiFast SYBR Green RT-PCR Kit (400) $944.00 For 400 x 25 µl reactions: 3 x 1.7 ml 2x QuantiFast SYBR Green RT-PCR Master Mix (contains ROX dye), 100 µl QuantiFast RT Mix, 2 x 2 ml RNase-Free Water |
Cat No./ID:204156 QuantiFast SYBR Green RT-PCR Kit (2000) $3,898.00 For 2000 x 25 µl reactions: 25 ml 2x QuantiFast SYBR Green RT-PCR Master Mix (contains ROX dye), 0.5 ml QuantiFast RT Mix, 20 ml RNase-Free Water |
Product Details
The QuantiFast SYBR Green RT-PCR Kit delivers highly specific and sensitive results, outperforming other real-time RT-PCR kits used in fast cycling mode (see figure "Specific one-step RT-PCR"). RT-PCRrun times are reducedby up to 60% (see figure "Significantly reduced PCR times"), allowing you to get results faster. You can also greatly increase your sample throughput or efficiently share a cycler with other users.
Results in one-step RT-PCR are comparable to those achieved in two-step RT-PCR (see table).
| Mean CT value | ||
|---|---|---|
| Amount of cDNA/RNA | QuantiFast SYBR Green PCR Kit (two-step RT-PCR) | QuantiFast SYBR Green RT-PCR Kit (one-step RT-PCR) |
| 10 ng | 20.27 | 20.11 |
| 1 ng | 23.44 | 23.46 |
| 0.1 ng | 27.18 | 26.86 |
| NTC | 45.00 | 45.00 |
QuantiFast SYBR Green RT-PCR Kit enables accurate quantification of targets over several log dilutions of template (see figure "Detection over 7 logs of template"). Even small differences in the amount of low-copy targets can be clearly distinguished.
The QuantiFast SYBR Green RT-PCR Kit delivers highly sensitive and specific results over a wide dynamic range on both standard and fast cyclers. The fluorescent dye SYBR Green I in the master mix enables the analysis of many different targets without having to synthesize target-specific labeled probes. A specially developed fast RT-PCR buffer contains the novel PCR additive Q-Bond, which significantly reduces denaturation, annealing, and extension times (see figure "Fast primer annealing"). A balanced combination of K+ and NH4+ ions in the PCR buffer promotes specific primer annealing, enabling 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. The optimized QuantiFast RT Mix enables cDNA synthesis in just 10 minutes.
| Component | Features and benefits | Benefits |
|---|---|---|
| HotStarTaq Plus DNA Polymerase | 5 min activation at 95ºC | Set up of qPCR reactions at room temperature |
| QuantiFast SYBR Green RT-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 | |
| SYBR Green I dye | Yields a strong fluorescent signal upon binding to double-stranded DNA | Highly sensitive quantification |
| 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 |
| QuantiFast RT Mix | Special blend of reverse transcriptases with high affinity for RNA | RNA can be transcribed in just 10 minutes, even through complex secondary structures |
The QuantiFast SYBR Green RT-PCR Kit is a ready-to-use master mix that eliminates the need for optimization of reaction and cycling conditions. Simply add primers and template to the ready-to-use PCR master mix, and start the reaction. Follow the protocol in the handbook to get fast and reliable results on any real-time cycler.
For optimal results in real-time, one-step RT-PCR, we recommend using the QuantiFast SYBR Green RT-PCR Kit in combination with QuantiTect Primer Assays. They are bioinformatically validated primer sets for any gene from human, mouse, rat, and many other species. Assays can be easily ordered online at the GeneGlobe Web portal.
The QuantiFast SYBR Green RT-PCR Kit is for use in gene expression analysis of RNA targets. QuantiFast SYBR Green RT-PCR Kits are compatible with all available real-time cyclers, including 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 SYBR Green RT-PCR Kit, which has been specially developed for fast cycling on these instruments.
Specifications
Features | Specifications |
| Applications | SYBR Green-based, real-time, one-step RT-PCR |
| Reaction type | Real-time and one-step RT-PCR |
| Real-time or endpoint | Real-time |
| Sample/target type | RNA |
| Single or multiplex | Single |
| SYBR Green I or sequence-specific probes | SYBR Green I |
| Thermal cycler | All real-time cyclers (e.g. LC, RG, ABI) |
| With or without ROX | With ROX |
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