Applied Biosystems™ ViralSEQ™ Quantitative Sf-Rhabdovirus Kits

The Applied Biosystems ViralSEQ Quantitative Sf-Rhabdovirus Kit is a two-step RT-qPCR assay for the detection of Sf-rhabdovirus in recombinant products manufactured in insect cell lines using baculovirus expression systems.

Supplier:  Applied Biosystems™ A50496

Catalog No. A50496

Description

The Applied Biosystems ViralSEQ Quantitative Sf-Rhabdovirus Kit is a two-step RT-qPCR assay for the detection of Sf-rhabdovirus in recombinant products manufactured in insect cell lines using baculovirus expression systems. Sf-rhabdovirus is an adventitious virus that was identified in the Sf9 and Sf21 insect cell lines that are commonly used as host cells for the production of recombinant proteins, adeno-associated virus (AAV) vectors for gene therapy, and virus-like particles (VLPs). The assay can be used to quantify Sf-rhabdovirus in cell lines, as well as crude and purified products.

ViralSEQ Quantitative Sf-Rhabdovirus Kit is a two-step RT-qPCR assay for the detection of Sf-rhabdovirus in Sf9/Sf21 cell lines and products such as recombinant proteins, AAV, and VLPs manufactured using baculovirus expression systems. 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.

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
• Includes master mix, RT-primer, TaqMan Assay mix, and RNA standard
•Configuration with PrepSEQ kit available for optimized RNA recovery

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. A50485), 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.

Specifications

• Content And Storage: Sf-Rhabdovirus RNA PCR Reagents
  • 10X Assay Mix, 275 μL, protect from light
  • 25X RT-PCR Enzyme Mix, 110 μL
  • 2X 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 2−8°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.
  
  
• PCR Method: qPCR
• Quantity: 100 reactions
• Target: Sf-rhabdovirus
• Includes: ViralSEQ™ Quantitative Sf-Rhabdovirus Kit, PrepSEQ™ Nucleic Acid Sample Prep Kit

• Detection Method: Primer-probe
• Product Line: ViralSEQ™
• Shipping Condition: Dry Ice
• No. of Reactions: Assay: 100 Reactions, Sample Prep: 100 Reactions