QIAcuity High Multiplex Probe PCR Kit

• For multiplexing beyond 5-plex including amplitude multiplexing of up to 12-plex
• 4x concentrated master mix that enables loading of more sample
• Optimized for microfluidic use in QIAcuity Nanoplates
• REACH-compliant kit

The QIAcuity High Multiplex Probe PCR Kit contains a 4x concentrated, ready-to-use master mix optimized for microfluidic use in the QIAcuity Nanoplates. The kit enhances the specificity and efficiency of probe-based digital PCR to provide accurate singleplex, or up to 12-plex, analysis. You can set up 8-plex reactions by using six QIAcuity channels (one assay/channel) plus two Long Stokes-Shift (LSS) dyes. For analysis of up to 12 targets, you can perform amplitude multiplexing by mixing two different assays per channel. 

The QIAcuity High Multiplex Probe Master Mix is optimized to increase specificity for accurate quantification of gDNA or cDNA on the QIAcuity dPCR instruments.

The kit works in conjunction with the QIAcuity Digital PCR System and the QIAcuity Nanoplates.

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The QIAcuity digital PCR platform overcomes the challenges of high multiplexed PCR in part thanks to the custom crosstalk matrix in QIAcuity Software 3.0. This new software feature compensates for optical crosstalk, or signal overlap, between neighboring channels. Together with the specialized chemistry of the QIAcuity High Multiplex Probe PCR Kit, users can detect targets in up to eight channels: six standard optical channels and two hybrid channels that make use of hydrolysis probes with Long Stokes-Shift (LSS) dyes ( Figure 1).  

QIAcuity Software 3.1 and the QIAcuity High Multiplex Probe PCR Kit allow users to detect up to 12 targets in six channels ( Figure 2 and  Figure 3).

Thanks to a novel, antibody-mediated, hot-start mechanism, the QIAcuity High Multiplex Probe PCR Kit delivers singleplex or multiplex cDNA or gDNA analysis with the highest specificity. 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 PCR amplification.

The principle of the dPCR reaction in the nanoplates is described here.

The QIAcuity High Multiplex Probe Kit contains a ready-to-use 4x PCR master mix, eliminating the need to optimize reaction and cycling conditions. Simply add the template DNA and primer-probe sets to the master mix and follow the procedure outlined in the Quick-Start Protocol to get fast and reliable results on the QIAcuity.

The system integrates partitioning, thermocycling and imaging into a single fully automated instrument that takes users from the sample to result in under 2 hours. Analyze your results on the Software Suite, where you can find the concentration of your target sequence in copies per microliter and obtain data on positive samples or NTC for quality control purposes. This analysis can be extended to remote computers within the same local area network (LAN) for added convenience.

The QIAcuity High Multiplex Probe PCR Kit, in combination with the QIAcuity Digital PCR System and the QIAcuity Nanoplates, enables quantitative analysis of cDNA targets and gDNA for use in applications, including:

• Rare mutation detection
• Copy number variation analysis
• Microbial research including wastewater monitoring
• Pathogen detection
• Genotyping
• Gene expression analysis
• miRNA research