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RealHelix™ qRT-PCR Kit [v6] [UDG System] is a probe-based qRT-PCR reagent that efficiently facilitates reproducible, reliable detection of up to five RNA targets by fast multiplex in a single tube reaction (FAST: 45 min/40 cycles or ULTRAFAST: 25 min/40 cycles). The Enzyme Mix is an optimized blend of reverse transcriptase, antibody-coupled hot-start Taq polymerase, RNase inhibitor protein, and a heat-labile UDG. The 5x Reaction Buffer contains all required components, including Mg²⁺, dUTP, and dNTPs. These components help ensure sensitivity, accuracy, and specificity in addition to fast PCR performance. The applied UDG system prevents the carryover contamination of products from previous reactions.

Figure 1. Multiplex qRT-PCR using the RealHelix™ qRT-PCR Kit [v6] [UDG System].

10-fold dilutions of purified SARS-CoV-2 viral RNA (from 10^-4 to 10^-7) spiked into a swab sample were analyzed using three different sets of target-specific primers and probes (N gene, ORF8, and S gene). The amount of viral RNA used in the 10^-7 dilution fold corresponds to approximately 10² copies. The probes were labeled with Texas Red (N gene), JOE (ORF8), FAM (S gene). The multiplex reaction was tested using the RealHelix™ qRT-PCR Kit [v6] [UDG System] on a real-time PCR instrument, Bio-Rad CFX96.

Figure 2. Multiplex qRT-PCR using the RealHelix™ qRT-PCR Kit [v6] [UDG System].

Plasmid clone mixtures harboring the partial region for each SARS-CoV-2 were used as templates in this reaction. SARS-CoV-2 clone mixture (10⁶, 10⁵, 10⁴, 10³, 10² copies) were analyzed using three different sets of target-specific primers and probes (N gene, ORF8 and S gene). The probes were labeled with Texas Red (N gene), JOE (ORF8), and FAM (S gene). The multiplex reaction was tested on a real-time PCR instrument, Bio-Rad CFX96.

Storage

• Store below -20 ℃

Shelf life

• 12 months