For SYBR Green-based and probe-based, real-time quantitative PCR for microbiome or quality control applications using reagents from Ultra Clean Production

Features

Nucleic acid-depleted reagents prevent background from residual DNA

Visual pipetting control prevents errors in pipetting

Exceptional inhibitor robustness for reliable results from all common starting materials

Convenient and automatable due to room-temperature reaction setup

Guard-protected, antibody-mediated hot-start mechanism for superior specificity

Product Details

The UCP SYBR Green PCR Kit provides a convenient, ultra-clean master mix format for highly sensitive and specific real-time PCR using any DNA or cDNA template. Dedicated processes are implemented to enable Ultra Clean Production of the master mix which minimizes contaminating DNA background. The master mix also comes with an in-built color tracker for identifying correct pipetting. Superior inhibitor resistance and a quantification option make the UCP SYBR Green PCR Kit an ideal choice for contamination-free, microbial applications, such as 16S sequencing (bacteria) or 18S sequencing (fungal) analysis. The kit is also highly recommended for quality control applications.

Combining superior performance with Ultra Clean Production, the UCP Probe PCR Kit is a unique solution for probe-based real-time PCR, free from residual DNA background. The Ultra Clean Production master mix is particularly suited for those applications which call for reagents depleted of contaminating nucleic acids such as quality control or microbiome analysis. Other attributes include a unique antibody-mediated hot-start mechanism, visual indicator for pipetting control, room temperature reaction setup and high resistance to inhibitors.

Performance

The nucleic acid-depleted, ultra-clean reagents of UCP PCR Kits are uniquely suited for processes that are prone to contaminating background DNA. Our Ultra Clean Production processes minimize potential contamination with residual DNA from expression host or other sources. All kit components undergo rigorous testing for bacterial and fungal DNA using generic PCR assays for 16S and 18S rRNA coding sequences. The kits are 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 kits also allow quantification over a wide linear range down to two E.coli bacterial genome (16S) or five C. albicans genome (18S) equivalents due to an optimized master mix formulation. Other highlights of UCP PCR Kits 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.

Principle

UCP PCR Kits deliver 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 . 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.

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 .

Procedure

UCP PCR Kits contain 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 deliver your 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.

Applications

UCP PCR Kits are 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.