QIAcuity EG PCR Kit (1mL)

From QIAGEN, the QIAcuity EG PCR Kit provides a 3x concentrated, ready-to-use EvaGreen PCR Mastermix optimized for microfluidic use in QIAcuity Nanoplates. The kit is designed for quantification of DNA or cDNA targets using the dsDNA-binding dye EvaGreen in singleplex reactions on QIAcuity digital PCR instruments. EvaGreen provides a higher fluorescence signal than SYBR Green at the same concentrations and delivers maximum amplification efficiency, specificity and sensitivity in dPCR. The mastermix contains an optimized fluorescent reference dye for reliable detection of proper partition filling in dPCR Nanoplates.

The mastermix incorporates QuantiNova DNA Polymerase with a novel antibody-mediated hot-start mechanism. At low temperatures, the QuantiNova DNA Polymerase is kept inactive by the QuantiNova Antibody together with QuantiNova Guard, which stabilizes the complex and prevents extension of nonspecifically annealed primers and primer–dimers. Within 2 minutes at 95°C, the antibody and guard are denatured and the polymerase is activated. The PCR initial heat activation step is 2 min at 95°C, followed by 3-step cycling for 40 cycles with denaturation at 95°C for 15 s, annealing at 55–62°C for 15 s and extension at 72°C for 15 s, with a final cooling step of 5 min at 40°C. For optimal efficiency, amplicons should ideally be 60–150 bp in length.

The QIAcuity PCR mixes can be stored at 30°C for up to 100 hours without impairing the performance of subsequent reactions, making the kit suitable for high-throughput reaction setup and plate-stack handling. Upon receipt, the kit should be stored at –30 to –15°C in a constant-temperature freezer and protected from light; the mastermix can alternatively be stored protected from light at 2–8°C. Under these conditions, the components are stable for 6 months without showing any reduction in performance. The system integrates partitioning, thermocycling and imaging into a single fully automated instrument, taking users from sample to result in under 2 hours, with analysis providing concentration in copies per microliter of the target sequence.