QuantiNova SYBR Green PCR Kit (2500)

Designed for SYBR Green-based real-time PCR, this QIAGEN kit delivers enhanced specificity through a novel antibody-mediated hot-start mechanism. At low temperatures, the QuantiNova Guard additive stabilises a binding complex between QuantiNova DNA Polymerase and the QuantiNova Antibody, keeping the polymerase inactive. Within two minutes of raising the temperature to 95°C, the complex is disrupted and the DNA polymerase becomes active, preventing extension of non-specifically annealed primers and the formation of primer-dimers. The chemistry supports ultrafast cycling for increased throughput, enables robust detection down to a single target copy, and produces accurate quantitative results from cDNA or gDNA on any real-time PCR cycler.

A built-in visual pipetting control helps to ensure accurate reaction setup. The master mix contains an inert blue dye, while the QuantiNova Yellow Template Dilution Buffer contains an inert yellow dye; once the diluted template is added to the master mix, the colour of the solution changes from blue to green, providing immediate visual confirmation that each reaction was assembled correctly. The dilution buffer is optional and does not restrict the user to a particular template buffer. Performance is reported to be seamless when the kit is combined with QuantiTect Primer Assays, and amplification and quantification are described as robust and sensitive across real-time PCR instruments from multiple manufacturers, with a ROX Reference Dye supplied for instruments that require a passive reference.

Quantification is precise and consistent over a dynamic range of eight orders of magnitude, with the underlying data demonstrating accurate analysis of HeLa cDNA from 100 ng down to 10 fg using an in-house beta-actin assay on the Applied Biosystems ViiA 7 and Bio-Rad CFX96. By combining the stringent hot-start with proven buffer technology, the chemistry is intended to deliver real-time PCR success at the first attempt and to fit easily into existing workflows.