Rotor-Gene SYBR® Green RT-PCR Kit
For ultrafast, one-step qRT-PCR gene expression analysis using SYBR Green I on Rotor-Gene cyclers
The Rotor-Gene SYBR Green RT-PCR Kit is designed for use with the Rotor-Gene Q and other Rotor-Gene cyclers, providing ultrafast, highly specific quantification of RNA targets with real-time one-step RT-PCR using SYBR Green I detection. Outstanding performance is achieved through the combination of a specially optimized master mix and the unique Rotor-Gene cycler. For convenience, the master mix can be stored at 2–8°C.
The Rotor-Gene SYBR® Green RT-PCR Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
Performance
When the Rotor-Gene SYBR Green RT-PCR Kit is used together with QuantiTect Primer Assays, highly sensitive quantification of specific PCR products is achieved without the need for optimization (see figures "Specific detection without the need for optimization").
Principle
The Rotor-Gene SYBR Green RT-PCR Kit enables reliable real-time RT-PCR quantification on the Rotor-Gene Q without the need for optimization of reaction and cycling conditions. Real-time one-step RT-PCR is carried out, which means that RNA is used as template in a reaction where reverse transcription and PCR take place sequentially in the same reaction vessel. Since it is not necessary to transfer the finished RT reaction to another tube for PCR, the real-time RT-PCR procedure is streamlined, making high-throughput analysis possible. 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. Highly specific amplification is assured through a balanced combination of K+ and NH4+ ions, which promote specific primer annealing, enabling high PCR specificity and sensitivity (see figure "Specific primer annealing"). Fast cycling without compromising performance is achieved using Q-Bond, a novel PCR additive that considerably shortens cycler run times (see figure "Fast primer annealing").
Procedure
A ready-to-use master mix eliminates the need for optimization of reaction and cycling conditions. Simply add template RNA, primers, and the supplied reverse transcriptase mix to the master mix and program the cycler. Instructions are provided in the detailed handbook supplied with the kit. For gene expression analysis using real-time one-step RT-PCR, the combination of the Rotor-Gene SYBR Green RT-PCR Kit with QuantiTect Primer Assays and the Rotor-Gene Q provides a complete, ready-to-run solution. QuantiTect Primer Assays 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.
Applications
The Rotor-Gene SYBR Green RT-PCR Kit provides rapid real-time quantification of RNA targets on the Rotor-Gene Q. The kits are also compatible with the Rotor-Gene 3000 and the Rotor-Gene 6000.
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