Rotor-Gene Multiplex PCR Kit
For ultrafast, multiplex, real-time PCR and two-step RT-PCR on Rotor-Gene cyclers
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. Buy Products
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.
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. Specifications
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