QuantiNova Probe PCR Kit
For highly sensitive, specific, and ultrafast, probe-based real-time PCR
Avoid the deleterious effects of mispriming at lower temperatures with the QuantiNova Probe PCR Kit. Based on a novel, antibody-mediated, hot-start mechanism, the kit enhances the specificity and efficiency of probe-based real-time PCR to provide accurate, singleplex or duplex, cDNA or gDNA analysis on various real-time PCR cyclers. The added convenience of extreme stability for up to 100 hours at room temperature, without a need for any cooling agent, makes it ideal for handling of high-throughput samples and automated workflows. An in-built tracking system for visual identification of correct pipetting gives you absolute peace of mind while the extreme sensitivity for even low target amounts ensures reliable qPCR results every time.
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Built-in visual control for correct pipettingThe master mix supplied with the QuantiNova Probe 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.
Reaction stability of up to 100 hoursThe real-time PCR mix can be stored at 30°C for up to 100 hours without impairing the performance of subsequent reactions (Table 1). The outstanding stability, even after extended storage at room temperature without the use of any cooling agent, makes the QuantiNova Probe PCR Kit ideal for high-throughput reaction setup and plate-stack handling (see table "Effect of storage on reaction stability").
Accurate and robust detection of a single DNA moleculeThe high sensitivity of the QuantiNova Probe PCR Kit results in accurate and robust detection of even a single target copy (see figure Sensitive and robust detection of a single target copy). The special master mix supplied with these kits enables accurate quantification of 2 targets having widely differing abundance in a single tube. This saves time, money, and reduces the amount of sample material needed. Moreover, the duplex PCR data obtained is comparable with that obtained from a singleplex PCR (see figure Comparable results in duplex and singleplex PCR).
Superior performance for difficult targetsThe unique combination of QIAGEN's proprietary and well-proven buffer technology along with the new QuantiNova DNA Polymerase, QuantiNova Antibody, and QuantiNova Guard, ensures real-time PCR success at the first attempt, without the need for costly and time-consuming optimization, even with challenging real-time PCR assays (see figure Superior results with challenging assays).
Highly consistent results on all real-time cyclersThe QuantiNova Probe PCR Kit can be used on any real-time cycler. ROX is provided in a separate tube and can be added if using a cycler that uses ROX as a passive reference dye. The kit delivers highly consistent results among different cyclers in terms of sensitivity, reproducibility, and efficiency. The consistency of the results is maintained, despite varying fast-cycling capacities of the different cyclers that result in different cycling protocols (see figure Consistent results with various real-time PCR cyclers).
The QuantiNova Probe PCR Kit delivers singleplex or duplex, cDNA or gDNA analysis with highest specificity because of a novel, antibody-mediated, hot-start mechanism (see figure Novel antibody mediated hot-start mechanism). At low temperatures, the QuantiNova DNA Polymerase is kept in an inactive state by the QuantiNova Antibody and a novel additive, QuantiNova Guard, that stabilizes the complex. This improves the stringency of the hot-start and prevents extension of nonspecifically annealed primers and primer–dimers. Within 2 minutes of raising the temperature to 95°C, the QuantiNova Antibody and QuantiNova Guard are denatured and the QuantiNova DNA Polymerase is activated enabling the PCR amplification.
QuantiNova Probe PCR Kits contain ready-to-use master mixes that eliminate the need for optimization of reaction and cycling conditions. Simply add template gDNA or cDNA, primers, and probe to the master mix and follow the protocol in the handbook to get fast and reliable results on any real-time cycler. ROX passive reference dye is provided as a separate tube and the concentrations of the dye can be adjusted depending on the type of cycler used, enabling use of the QuantiNova Probe PCR kits on virtually any real-time cycler. Due to the optimized ROX concentrations, detection of even low copy numbers is achieved through automatic data analysis.
QuantiNova Probe PCR Kits can be used for probe-based gene expression analysis of cDNA targets and quantitative gDNA analysis on any real-time cycler. This includes instruments from Applied Biosystems, Bio-Rad, Cepheid, Eppendorf, Roche, and Agilent.
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