The Applied Biosystems ViralSEQ Quantitative Sf-Rhabdovirus Kit is a two-step RT-qPCR assay for the detection of sf-rhabdovirus from the Sf9/Sf21 baculovirus platform used in the development of gene therapies, cell-based vaccines, and similar biotherapeutics. This assay leverages a tagged RT-primer design containing a non-viral sequence to enable specific and accurate quantification of negative-sense genomic sf-rhabdovirus RNA. This allows for a more accurate quantification of true viral count and avoids the overestimation that is usually caused by mRNA in non-strand-specifc assay designs.
This kit is bundled together with sample preparation reagents in the form of a PrepSEQ Nucleic Acid Sample Preparation Kit (Cat. No. A50485), which is optimized for highly efficient RNA recovery from common bioproduction matrics.
Features of the ViralSEQ Quantitative Sf-Rhabdovirus Kit include: • Negative-sense-specific RT-PCR assay design for accurate quantification of Sf-rhabdovirus • Specific to genomic Sf-rhabdovirus RNA; not impacted by the presence of positive-strand mRNA; no cross-reactivity with unrelated DNA/RNA • Manual and automated sample preparation with PrepSEQ kit, optimized for quantitative recovery of RNA from common bioproduction sample matrices • Includes master mix, RT-primer, TaqMan Assay mix, and RNA standard
Integrated workflow solution The ViralSEQ Quantitative Sf-Rhabdovirus Kit is part of an integrated workflow for impurity and contaminant testing during biopharmaceutical manufacturing. The kit is compatible with the PrepSEQ Nucleic Acid Sample Preparation Kit (Cat. No. A50484) (included), which offers both a manual and automated sample preparation workflow. The RT step is performed on the Veriti 96-Well Fast Thermal Cycler ( 0.1-mL block configuration). For real time PCR, the ViralSEQ kit has been validated on the Applied Biosystems 7500 Fast Real-Time PCR System and the Applied Biosystems QuantStudio 5 Real-Time PCR System. Data analysis is streamlined using AccuSEQ Real-Time PCR Detection Software that provides accurate quantitation and security, audit, and e-signature capabilities to help enable 21 CFR Pt 11 compliance.
Contents & Storage
Sf-Rhabdovirus RNA PCR Reagents • 10 x Assay Mix, 275 µL, protect from light • 25 x RT-PCR Enzyme Mix, 110 µL • 2 x RT-PCR Buffer, 1.375 mL • RT Primer, 55 µL • Nuclease Free Water, 1.0 mL, store at -15°C to -25°C until first use. After first use, store at 2°C to 8°C. Store all at -15°C to -25°C unless otherwise indicated.
Sf-Rhabdovirus RNA Control • Sf-rhabdovirus RNA Control (1.25 x 108 copies/µL), 44 µL • RNA Dilution Buffer, 7 mL Store all at -15°C to -25°C.
PrepSEQ Nucleic Acid Extraction Kit • Lysis Buffer (2 Bottles, 50 mL/bottle) • Binding Solution/Isopropanol (1 empty bottle) • Wash Buffer Concentrate (2 Bottles, 26 mL/bottle) • Elution Buffer (1 Bottle, 25 mL) • Proteinase K Buffer(1 Bottle, 50 mL) • Magnetic Particles (2 Tubes, 1.5 mL/tube), store at 2°C to 8°C • Proteinase K (20 mg/mL) (1 Tube, 1.25 mL), store at or below -20°C • Pk. Buffer II (1 Bottle, 11 mL) Store all at room temperature unless otherwise indicated.
PrepSEQ REsidual DNA Sample Preparation Kit • Proteinase K (20 mg/mL) (1 Tube, 1.25 mL) • Yeast tRNA (10 mg/mL) (1 Tube, 0.5 mL) • Glycogen (5 mg/mL) (2 Tubes, 1.0 mL/tube) Store all at or below -20°C.