UCP SYBR^® Green PCR Kit

The nucleic acid-depleted, ultra-clean master mix of the UCP SYBR Green PCR Kit is uniquely suited for those processes which suffer from contaminating background DNA. Our Ultra Clean Production processes minimize potential contamination with residual DNA from expression host or other sources. All components of the kit undergo rigorous testing for bacterial and fungal DNA using generic PCR assays for 16S and 18S rRNA coding sequences. This makes the UCP SYBR Green PCR Kit your go-to kit for microbial testing, quality control procedures or microbiome/metagenome analysis and sequencing. Testing low biomass samples will also benefit from ultra-clean reagents. The kit’s performance is further enhanced by improved inhibitor resistance and minimal GC-content which ensure accurate results. Optimized master mix formulation allows quantification over a wide linear range – 2 E.coli bacterial genome (16S) or 5 C. albicans genome (18S) equivalents. Integrated visual pipetting, room-temperature setup and ultrafast cycling further improve efficiency in microbial workflows.

The UCP SYBR Green PCR Kit delivers high specificity using an efficient 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 stringent hot-start mechanism prevents any enzymatic activity at ambient temperatures allowing reaction setup without ice or on instruments. The enzyme is fully activated after the 2-minute incubation step at 95°C and starts amplifying with high specificity from the first cycle. The hot-start procedure also prevents extension from non-specifically annealed primers and primer-dimers in the first cycle ensuring a highly specific and reproducible PCR.

The built-in color pipetting feature ensures accurate reaction setup. The master mix contains an inert blue dye whereas the UCP Yellow Template Dilution Buffer contains an inert yellow dye. When the diluted template nucleic acid 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 SYBR Green PCR Kit provides a convenient ready-to-use 2x concentrated master mix formulation, 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 and delivers highly consistent results with different cyclers in terms of sensitivity, reproducibility and efficiency, despite varying fast-cycling capacities. UCP ROX is provided in a separate tube and can be added if using a cycler that requires ROX as a passive reference dye.

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

The UCP SYBR Green PCR Kit is ideally suited for procedures demanding high purity and accuracy, such as 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. It 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 any 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.