QuantiTect SYBR® Green RT-PCR Kit
For one-step qRT-PCR using SYBR Green I for gene expression analysis
- High specificity in one-step RT-PCR
- Reliable quantification of low-abundance transcripts
- Accurate quantification over several logs of template
- No need to optimize reaction and cycling conditions
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- Product Details
- Specifications
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Cat No./ID:204243 QuantiTect SYBR Green RT-PCR Kit (200) $932.00 For 200 x 50 µl reactions: 3 x 1.7 ml 2x QuantiTect SYBR Green RT-PCR Master Mix, 100 µl QuantiTect RT Mix, 2 x 2 ml RNase-Free Water |
Cat No./ID:204245 QuantiTect SYBR Green RT-PCR Kit (1000) $3,767.00 For 1000 x 50 µl reactions: 25 ml 2x QuantiTect SYBR Green RT-PCR Master Mix, 0.5 ml QuantiTect RT Mix, 20ml RNase-Free Water |
Product Details
QuantiTect SYBR Green RT-PCR Kits contain an optimized, ready-to-use master mix for highly specific and sensitive real-time quantification of RNA targets using SYBR Green I. 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 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, an optimized mix of reverse transcriptases enables cDNA synthesis from a wide range of RNA template amounts, while HotStarTaq DNA Polymerase provides a stringent hot start, preventing the formation of nonspecific products.
QuantiTect SYBR Green 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 min activation at 95ºC | Set-up of qPCR reactions at room temperature |
| QuantiTect SYBR Green 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 | Eliminates contamination from carryover of PCR products by optional UNG treatment |
| SYBR Green I dye | Yields a strong fluorescent signal upon binding double-stranded DNA | Highly sensitive quantification |
| 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 SYBR Green RT-PCR Kit overcomes the need for optimization of reaction conditions, which can be tedious and time-consuming. Simply add primers and 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.
Highly specific results in gene expression analysis are guaranteed when QuantiTect SYBR Green RT-PCR Kits are used in combination with QuantiTect Primer Assays. These are genomewide, bioinformatically validated primer sets for detecting transcripts from human, mouse, rat, and many other species. QuantiTect Primer Assays can be easily ordered online at GeneGlobe.
The QuantiTect SYBR Green RT-PCR Kit is for use in gene expression analysis of RNA targets. QuantiTect 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 | Real-time quantification of RNA targets |
| 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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