QuantiNova Multiplex PCR Kits
For ultrafast, multiplex, real-time PCR and two-step qRT-PCR using sequence-specific probes
QuantiNova Multiplex PCR Kits enable fast and reliable quantification of up to 5 cDNA or gDNA targets in a single tube by multiplex, real-time PCR or two-step RT-PCR. Q-bond technology and an optimized master mix promote ultrafast multiplex real-time PCR within 1 hour. The combination of a unique hot start and PCR buffer system in the ready-to-use 4x master mix ensures highly sensitive qPCR on any real-time cycler without the need for optimization, also providing automated reaction set up options at room temperature.
An in-built tracking system for visual identification of correct pipetting prevents human errors, and the combination with QuantiNova Reverse Transcription Kit for cDNA synthesis allows inclusion of QuantiNova Internal Control RNA to monitor successful reverse transcription and qPCR.
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The special master mix supplied with QuantiNova Multiplex PCR Kits allows rapid setup of multiplex reactions and delivers successful results at the first attempt, providing multiplex PCR data that are comparable with singleplex PCR data. The highly concentrated 4x master mix accommodates up to 800ng template input ensuring outstanding sensitivity, even in up to 5-plex reactions. The kit can clearly distinguish between small differences in the amount of template and provides accurate quantification of targets of widely differing abundance.
A novel, antibody-mediated, hot-start mechanism (see figure Novel antibody mediated hot-start mechanism) ensures outstanding specificity and allows reaction set up at room temperature, ideally suited for automated procedures. The specially developed fast PCR buffer contains the additive Q-Bond, which significantly reduces annealing and extension times, allowing multiplex qPCR in less than one hour. The visual pipetting control prevents human errors (see figure Built-in pipetting control) and increases process safety, particularly if combined with the Internal Control RNA provided in the QuantiNova Reverse Transcription Kit for quantitative 2-step RT-PCR.
QuantiNova Multiplex PCR Kits deliver highly sensitive and rapid results over a wide dynamic range on both standard and fast cyclers without optimization. The specially developed fast PCR buffer contains the additive Q-Bond, which significantly reduces annealing and extension times (see figure "Fast primer annealing").
Amplifying reference and target genes in the same reaction instead of in separate reactions increases the reliability of gene quantification by minimizing handling errors. Additionally the Internal Control RNA provided in the QuantiNova Reverse Transcription Kit for quantitative 2-step RT-PCR can be incorporated to monitor successful reverse transcription and qPCR.
The master mix supplied with the QuantiNova Multiplex PCR Kit contains an inert blue dye that does not interfere with the real-time PCR, but increases visibility in the tube or well. The QuantiNova Yellow Template Dilution Buffer contains an inert yellow dye. When the template nucleic acid, diluted with the QuantiNova Yellow Template Dilution Buffer, is added to the master mix, the color of the solution changes from blue to green (see figure Built-in pipetting control), providing a visual indication of correct pipetting and reaction setup.
The QuantiNova Multiplex PCR Kit master mix can conveniently be stored at 2-8° C for up to 12 months, and also the reaction set up is extraordinary stable at room temperature, allowing automated procedures to increase efficiency and accuracy.
QuantiNova Multiplex PCR Kits contain ready-to-use 4x master mix that eliminate the need for optimization of reaction and cycling conditions. Simply add up to 800ng template DNA and primer-probe sets to the master mix and follow the protocol in the handbook to get fast and reliable results on any real-time cycler. Kits provide ROX passive reference dye in a separate tube, to adjust appropriate ROX concentration, if required for your instrument.
For optimal results in real-time two-step RT-PCR, we recommend synthesizing cDNA using the QuantiNova Reverse Transcription Kit, which provides fast cDNA synthesis in just 20 minutes with integrated removal of genomic DNA contamination. It additionally provides the QuantiNova Internal Control RNA, offering an in-process monitoring of successful reverse transcription and qPCR.
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