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UCP SYBR® Green PCR Kit

SYBR Green-based, real-time quantitative PCR using reagents from Ultra Clean Production
  • Nucleic acid-depleted reagents prevent background from residual DNA
  • Exceptional inhibitor robustness for reliable results from all common starting materials
  • Room-temperature reaction setup allows automation
  • Guard-protected hot-start chemistry provides superior specificity and sensitivity
  • Visual pipetting control dyes monitor successful procedure
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 microbiome applications like 16S or 18S analysis. The kit is also highly recommended for quality control applications.
Cat No./ID: 208012
UCP SYBR Green PCR Kit (100)

1 x 1 ml UCP SYBR Green PCR MasterMix (2x); 1 x 200 µl UCP Yellow Template Dilution Buffer; 1 x 250 µl UCP ROX Dye; 1 x 1.9 ml UCP PCR Water

Cat No./ID: 208014
UCP SYBR Green PCR Kit (500)
5 x 1 ml UCP SYBR Green PCR MasterMix (2x); 1 x 200 µl UCP Yellow Template Dilution Buffer; 2 x 250 µl UCP ROX Dye; 4 x 1.9 ml UCP PCR Water
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Built-in pipetting control
The master mix contains an inert blue dye. Combined with UCP Yellow Template Dilution Buffer, the resulting solution turns green, indicating that the reaction was set up correctly.
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Principle of Guard-protected hot-start mechanism
The DNA Polymerase is kept in an inactive state by the antibody and Guard molecule until the initial heat activation step.
Performance
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.
Principle

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).

Procedure

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.

Applications

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.

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