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Rotor-Gene Multiplex PCR Kit

For ultrafast, multiplex, real-time PCR and two-step RT-PCR on Rotor-Gene cyclers

  • Optimized for ultrafast, reliable results on Rotor-Gene cyclers
  • Sensitive detection of up to 4 targets in 1 tube
  • Successful multiplex PCR without the need for optimization
  • Reliable quantification of low- and high-abundance targets

The Rotor-Gene Multiplex PCR Kit is designed for use with the Rotor-Gene Q and other Rotor-Gene cyclers, providing ultrafast, highly reliable quantification in multiplex, real-time PCR and two-step RT-PCR using sequence-specific probes. Depending on the cycler configuration, up to 4 cDNA or gDNA targets (e.g., 1 control gene and 3 target genes) can be quantified simultaneously in the same tube. 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.

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Cat No./ID: 204772
Rotor-Gene Multiplex PCR Kit (80)
Trial kit for 80 x 25 µl reactions: 1 ml 2x Rotor-Gene Multiplex PCR Master Mix, 2 ml RNase-Free Water
Cat No./ID: 204774
Rotor-Gene Multiplex PCR Kit (400)
For 400 x 25 µl reactions: 3 x 1.7 ml 2x Rotor-Gene Multiplex PCR Master Mix, 2 x 2 ml RNase-Free Water
The Rotor-Gene Multiplex PCR Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.

Product Details

Performance
With the Rotor-Gene Multiplex PCR Kit, up to 4 cDNA targets can be simultaneously and rapidly quantified in the same tube, increasing throughput and saving precious sample material (see figures "Reliable multiplex analysis" and "Reliable duplex analysis"). Genes of different expression levels are all amplified in the same tube with the same high efficiency, enabling reliable relative quantification of gene expression.
Principle
Amplifying reference and target genes in the same reaction instead of in separate reactions increases the reliability of gene quantification by minimizing handling errors. The Rotor-Gene Multiplex PCR Kit enables reliable multiplex real-time PCR quantification of cDNA targets on the Rotor-Gene Q without the need for optimization of reaction and cycling conditions (see flowchart "QIAGEN multiplex kits"). Highly specific amplification is assured through a balanced combination of K+ and NH4+ ions, which promote specific primer annealing and enable high PCR specificity and sensitivity, while synthetic Factor MP, an innovative PCR additive specially developed for challenging multiplex PCR applications, allows different amplicons in the same reaction to all be amplified with the same high efficiency (see figure "Unique PCR buffer").

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"). In addition, the highly stringent hot-start enzyme HotStarTaq Plus DNA Polymerase is rapidly activated at the start of PCR by a brief 5-minute incubation at 95ºC. 

Components of 2x Rotor-Gene Multiplex PCR Kit*
ComponentFeatures Benefits
HotStarTaq Plus DNA Polymerase 5 min activation at 95ºC Set up of qPCR reactions at room temperature
Rotor-Gene Multiplex PCR Buffer Balanced combination of NH4+ and K+ ions Specific primer annealing ensures reliable qPCR results
Synthetic Factor MP Reliable multiplexing analysis of up to 4 targets in the same tube
Unique Q-Bond additive Faster PCR run times enable faster results and more reactions per day
* Also contains dNTP mix (dATP, dCTP, dGTP, dTTP).
Procedure

A ready-to-use master mix eliminates the need for optimization of reaction and cycling conditions. Simply add template cDNA and primer-probe sets to the master mix and program the cycler. The handbook supplied with the kit lists recommended dyes and contains a single protocol for all multiplex qPCR assays.

For optimal results in real-time two-step RT-PCR, we recommend synthesizing cDNA using the QuantiTect Reverse Transcription Kit. The kit provides fast cDNA synthesis in just 20 minutes with integrated removal of genomic DNA contamination.

Applications

The Rotor-Gene Multiplex PCR Kit is optimized for fast, real-time PCR and two-step RT-PCR analysis using sequence-specific probes on the Rotor-Gene Q. It is also compatible with the Rotor-Gene 3000 and the Rotor-Gene 6000. Up to 4 cDNA or gDNA targets can be simultaneously and rapidly quantified in the same tube, increasing throughput and saving precious sample material.

For ultrafast, one-step qRT-PCR multiplex analysis of RNA targets using sequence-specific probes on Rotor-Gene cyclers, use the Rotor-Gene Multiplex RT-PCR Kit.

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Kit Handbooks (1)
For fast multiplex real-time PCR, two-step RT-PCR, and one-step RT-PCR using sequence-specific probes on Rotor-Gene cyclers
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