UCP Probe PCR Kit

Here is a kit that gets rid of background signals from residual DNA in your qPCR. The UCP Probe PCR Kit’s nucleic acid-depleted reagents minimize potential contamination with residual DNA from the expression host or other sources. All components of the kit are meticulously tested for bacterial and fungal DNA using generic PCR assays for 16S and 18S rRNA coding sequences. The kit is highly recommended for applications that warrant ultra-pure reagents such as microbial testing, quality control procedures, microbiome/metagenome analysis and sequencing and low biomass sample testing. The kit also allows quantification over a wide linear range down to 2 E.coli bacterial genome (16S) or 5 C. albicans genome (18S) equivalents due to an optimized master mix formulation. Other highlights of the kit are superior robustness versus inhibitors or GC content variations, integrated visual pipetting, room-temperature setup and ultrafast cycling, all of which improve the efficiency of your microbial workflows.

High specificity of the UCP Probe PCR Kit can be attributed to an antibody-mediated, Guard-protected hot-start mechanism. At low temperatures, the DNA Polymerase is kept in an inactive state by an antibody and a novel Guard additive, that stabilizes the complex (see figure Principle of Guard-protected hot-start mechanism). This allows reaction setup without ice or on instruments as the enzyme is not active at ambient temperatures. The 2-minute incubation step at 95°C activates the enzyme, which starts amplifying with high specificity from the first cycle. The hot-start procedure also provides high specificity and reproducibility by preventing extension from non-specifically annealed primers and primer-dimers in the first cycle.

Two in-built inert dyes are used to provide a visual pipetting control. The UCP master mix contains an inert blue dye and the UCP Yellow Template Dilution Buffer contains an inert yellow dye. When the template nucleic acid, diluted with the UCP Yellow Template Dilution Buffer, is added to the master mix, the color of the solution changes from blue to green, providing a visual indication of correct pipetting and reaction setup (see figure Built-in pipetting control).

The UCP Probe PCR Kit contains a ready-to-use 2x concentrated master mix, UCP PCR water and UCP ROX dye for optional use. The ultra-fast two-step cycling protocols get you the results in less than an hour. The kit can be used on any real-time cycler. UCP ROX is provided in a separate tube and can be added if using a cycler that requires ROX as a passive reference dye. The kit delivers highly consistent results with different cyclers in terms of sensitivity, reproducibility and efficiency, despite varying fast-cycling capacities.

To further streamline PCR workflows, we recommend the QIAgility instrument for automated reaction setup.

The UCP Probe PCR Kit is ideally suited for procedures where contaminating nucleic acid background is often an issue. These include microbial testing, microbiome/metagenome workflows, quality control and other quantitative nucleic acid analyses. The kit offers high efficiency particularly in microbial analysis workflows such as quantification of microbial DNA versus host DNA, prior to NGS library preparation from swabs or body fluids. The kit is also highly recommended for detection and quantification of contaminating or residual DNA, such as bacterial, fungal or any host DNA in quality control procedures or biological production processes. The kit can be used on all common real-time cyclers.

For microbiome analysis, QIAGEN offers a range of dedicated products including DNeasy Power Soil Pro Kits for sample extraction, UCP Multiplex and HiFidelity Kits for end-point PCR, QIAseq 16S/ITS Panels for next-generation sequencing and CLC Microbial Genomics Pro Suite software for bioinformatic analysis.