Advanced reverse transcription performance
The new QIAcuity Advanced Reverse Transcription Mix includes QIAGEN´s new patent-pending HotStart reverse transcription technology. The new HotStart RT enzyme is non-functional at room temperature and prevents the generation of off-target cDNA even when left to incubate at 37°C for many hours (see figure Melt curve analysis of the new HotStart RT). It thereby enables a reaction setup at room temperature. The HotStart RT enzyme allows users to assemble their reactions at room temperature. Furthermore, and more importantly, the HotStart RT enzyme enables users to assemble and load multiple Nanoplates in parallel without needing to worry about unwanted and potentially unspecific RT activity while the plates wait at ambient temperatures. The HotStart RT enzyme is blocked while Nanoplates sit and wait inside the QIAcuity instrument until transferred to the thermal cycler, where the RT step of the protocol occurs.
Enhancer GC boosts specific assay performance
dPCR assay performance depends on the sequences and design of primer and probes. The GC content and amplicon length vary for industry catalog and custom-designed assays. PCR chemistries are optimized for universal usage and best performance; however, sometimes the addition of specific enhancers is recommended. The new Enhancer GC is an optional component (separate vial) you can add to the dPCR reaction when amplicons are GC-rich or are longer than 150 nt. It is also recommended for use with TaqMan (Thermo Fisher) gene expression assays.
Table: RT efficiency with and without Enhancer GC
||without Enhancer GC
||with Enhancer GC
|ABI TaqMan Assays
Superior amplification performance
The QIAcuity OneStep Advanced Probe Kit for hydrolysis probe-based dPCR uses the latest version of QIAGEN's high-quality DNA Polymerase. The unique combination of QIAGEN's proprietary and well-proven buffer technology optimized for the Nanoplate microfluidic along with the new QuantiNova DNA Polymerase delivers highly consistent results in terms of sensitivity, reproducibility and efficiency.