For ultrafast, real-time PCR and two step RT-PCR using SYBR Green on Rotor-Gene cyclers
Specific and sensitive detection of even low-copy targets
Accurate detection of a wide range of template amounts
Specially formulated master mix for reliable results without optimization
Guaranteed performance combined with QuantiTect Primer Assays
The Rotor-Gene SYBR Green PCR Kit is designed for use with the Rotor-Gene Q and other Rotor-Gene cyclers, providing ultrafast, highly specific quantification of gDNA and cDNA targets with quantitative real-time PCR or two-step RT-PCR using SYBR Green I detection. Outstanding qPCR 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.
For 2000 x 25 µl reactions: 25 ml 2x Rotor-Gene SYBR Green PCR Master Mix, 20 ml RNase-Free Water
Highly specific and sensitive detection.
Real-time two-step RT-PCR was carried out using tenfold dilutions of human leukocyte cDNA (10 ng to 0.1 ng) and a QuantiTect Primer Assay for BCL2 (B-cell CLL/lymphoma 2). Reactions were run in triplicate using either [A] the Rotor-Gene SYBR® Green PCR Kit and Rotor-Gene Q or [B] a kit and cycler from Supplier AII. The Rotor-Gene system provided lower CT values and greater reproducibility within triplicates. Melting curve analysis (see insets) showed a single peak, demonstrating specific amplification of the target.
Reliable and specific detection down to 10 copies.
Real-time PCR was carried out using tenfold dilutions of plasmid DNA (109 to 10 copies) and a QuantiTect Primer Assay for PPIA (cyclophilin A). Reactions were run in triplicate using the Rotor-Gene SYBR® Green PCR Kit and Rotor-Gene Q. [A] The Rotor-Gene system provided reliable detection over the entire range of template dilutions as well as highly reproducible CT values within each set of triplicates. [B] Melting curve analysis indicates specific amplification of the target.
When QuantiTect Primer Assays are used together with the Rotor-Gene SYBR Green PCR
Kit, highly sensitive quantification of specific PCR products is achieved without
the need for optimization (see table).
Superior performance in RT-PCR with SYBR Green
BAX (BCL2-associated X protein)
BCL2 (apoptosis gene)
b-Actin (housekeeping gene)
Human leukocyte cDNA (1 ng) was used as a template in SYBR Green-based real-time
two-step RT-PCR. Triplicate reactions were run using QuantiTect Primer Assays for
4 different targets: BAX, BCL2, MYC, and β-Actin.
The Rotor-Gene SYBR Green PCR Kit enables rapid and reliable real-time PCR quantification on the Rotor-Gene Q without the need for optimization of reaction and cycling conditions. 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 enables cycler run times of as low as 45 minutes (see figure "Fast primer annealing").
Components of 2x Rotor-Gene SYBR Green PCR Kit*
HotStarTaq Plus DNA Polymerase
5 min activation at 95ºC
Set up of qPCR reactions at room temperature
Rotor-Gene SYBR Green PCR Buffer
Balanced combination of NH4+ and K+ ions
Specific primer annealing ensures reliable qPCR results
Unique Q-Bond additive
Faster PCR run times enable faster results and more reactions per day
SYBR Green I dye
Yields a strong fluorescent signal upon binding double-stranded DNA
Highly sensitive quantification
* Also contains dNTP mix (dATP, dCTP, dGTP, dTTP).
The Rotor-Gene SYBR Green PCR Master Mix eliminates the need for optimization of reaction and cycling conditions. Simply add DNA template DNA and primers 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 two-step RT-PCR, the combination of the Rotor-Gene SYBR Green PCR Kit, QuantiTect Reverse Transcription Kit, QuantiTect Primer Assays, and Rotor-Gene Q provides a complete, ready-to-run solution. The QuantiTect Reverse Transcription Kit delivers fast cDNA synthesis in just 20 minutes with integrated removal of genomic DNA contamination. 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.
The Rotor-Gene SYBR Green PCR Kit provides rapid real-time quantification of cDNA and gDNA targets on the Rotor-Gene Q. The Kits are also compatible with the Rotor-Gene 3000 and the Rotor-Gene 6000.
For ultrafast, one-step qRT-PCR gene expression analysis of RNA targets using SYBR Green I on Rotor-Gene cyclers, use the Rotor-Gene SYBR Green RT-PCR Kit.
Real-Time quantification of genomic DNA or cDNA targets
For ultrafast quantitative real-time PCR and two-step RT-PCR using SYBR Green I on Rotor-Gene cyclers
For 100 x 50 µl reactions: QIAGEN OneStep RT-PCR Enzyme Mix (1 x 200 µl), 5x QIAGEN OneStep RT-PCR Buffer (1 x 1 ml), dNTP Mix (1 x 200 µl, 10 mM each), 5x Q-Solution (1 x 2 ml), RNase-Free Water (2 x 1.9 ml)