rcAAV-2/N Quantitation Kit SHENTEK®

For Research Use Only (RUO)

$1,751.00

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Sku: 1403444
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In stock & estimated to ship in 3-7 days by October 13, 2026
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Detection Method
qPCR
Assay Time
Refer to manual
Storage Temperature
-20 °C (stable for 24 months)

Product Description

SHENTEK® rcAAV Quantitation Kit is a qPCR-based kit for detection and quantitation of replication-competent adeno-associated virus (rcAAV) contamination in recombinant AAV (rAAV) serotypes designated as rAAV-2/N. The kit is designed to quantify rcAAV-2/N contamination in rAAV production samples including harvested cell-culture bulk, purified stocks and final products. The kit contains a T&R DNA control for preparation of an absolute quantitation standard curve, separate Target and Reference Primer & Probe mixes, qPCR reaction buffer and supporting reagents. The kit supports standard-curve (absolute) quantitation and is compatible with common qPCR platforms (e.g., ABI 7500, Bio-Rad CFX96, Roche 480, SHENTEK-96S).

Important: The ITR sequence of the rAAV test sample must match the provided ITR sequence for correct detection. Reagents supplied are sufficient for 2 × 100 reactions.

Technical Specifications

Parameter Details
Intended use Quantitative detection of replication-competent AAV (rcAAV-2/N) contamination in rAAV-2/N samples by qPCR (absolute quantitation, standard curve)
Number of reactions (kit size) Reagents for 2 × 100 reactions
Dynamic range / standard curve 20 to 2 × 10^6 copies/µL (standard curve concentrations: ST6=2×10^1 to ST1=2×10^6 copies/µL)
Limit of quantification (LOQ) 20 copies/µL (lowest standard used for quantitation)
Reaction volume 30 µL total reaction volume (10–10.04 µL qPCR mix + 20 µL sample/standard per well)
Thermal cycling (example ABI 7500) Pre-melt 95 °C 10:00 (1 cycle); Denature 95 °C 00:15; Anneal/extend 60 °C 00:30; Read at 72 °C 00:30; 40 cycles. (Note: ABI 7500 users set final read step to 72 °C 34 s.)
Detector channels / probes Target-2 detected in FAM channel; Reference-2 detected in CY5 channel; separate primer/probe mixes provided
Ct threshold settings (recommended) Reference-2 threshold = 0.06; Target-2 threshold = 0.02; use automatic baseline and manual Ct
Ct acceptance criteria NTC Ct (Reference and Target) should be >35 or undetermined. NCS Ct values should be larger than mean Ct of lowest standard.
Contamination calculation rcAAV contamination rate = (Detection value of Target genes) ÷ (Detection value of Reference genes × dilution factor ÷ 2). Note: each rAAV contains two copies of the Reference gene.
Compatible instruments ABI 7500, Bio-Rad CFX96, SHENTEK-96S, Roche 480 and other real-time PCR systems (ROX usage depends on instrument)
Storage stability Kit components can be stored under appropriate conditions for up to 24 months

Features

  • Designed for rcAAV-2/N detection: Specifically optimized to quantify replication-competent AAV contamination in rAAV-2/N serotypes using ITR-targeted assays.
  • Absolute quantitation by standard curve: Includes T&R DNA Control and dilution scheme to generate a 6-point standard curve (2×10^1 – 2×10^6 copies/µL) for absolute quantitation.
  • Complete assay components: Provides qPCR reaction buffer, Target and Reference primer/probe mixes, ROX option, DNA dilution buffer and DNA control for streamlined setup.
  • Platform flexibility: Compatible with multiple real-time PCR instruments; ROX addition is optional depending on instrument model.
  • Procedural controls and guidance: Includes NTC and NCS guidance, recommended plate layouts, Ct thresholds, and sample dilution recommendations to support robust QC workflows.

Applications

  • Quantitation of rcAAV contamination in recombinant AAV production (rAAV stocks and final products)
  • Quality control testing during rAAV manufacturing
  • Analysis of harvested cell-culture samples for presence of replication-competent AAV
  • Lot release and batch testing for rcAAV contamination

Kit Contents

T&R DNA Control-2 (lyophilized powder, 1 tube)-20 °C
rcAAV qPCR Reaction Buffer (400 µL × 4 tubes)-20 °C (protect from light)
Target Primer & Probe MIX-2 (200 µL × 1 tube)-20 °C (recommended)
Reference Primer & Probe MIX-2 (200 µL × 1 tube)-20 °C (recommended)
100× ROX (20 µL × 1 tube)-20 °C (recommended)
DNA Dilution Buffer (DDB) (1.5 mL × 4 tubes)-20 °C
ddH2O (1 mL × 1 tube)2–8 °C (or -20 °C long-term)
* Total: 2 × 100 reactions | Method: qPCR (Absolute Quantitation by Standard Curve)

Attention

• For Research Use Only (RUO). Not for use in diagnostic procedures.

• Read Material Safety Data Sheets (MSDS) and wear appropriate PPE (gloves, lab coat, eye protection, mask) when handling reagents.

• Prevent nucleic acid contamination: UV-treat surfaces, pipettes and tubes for 30 minutes and disinfect with 75% alcohol before setup.

• Benzonase nuclease treatment of rAAV stocks prior to extraction is required to remove non-encapsidated nucleic acids and improve direct-qPCR accuracy.

• Follow the SHENTEK Virus DNA & RNA Extraction Kit instructions for sample extraction when applicable.

• Thaw reagents at 2–8 °C or on ice, vortex and briefly centrifuge before use; protect light-sensitive reagents (reaction buffer) from light.

• Centrifuge the lyophilized DNA control at 12,000 rpm for 1 min and reconstitute precisely with 55 µL ddH2O before preparing dilutions.

• Prepare standard curve fresh (recommended six points) and run samples in triplicate for reliable quantitation.

• Add 100× ROX only if required by the qPCR instrument (ABI 7500 requires ROX; SHENTEK-96S and Roche 480 generally do not).

• Use recommended Ct threshold settings (Reference = 0.06; Target = 0.02) and check slope/intercept/R² of standard curve for assay validity.

Quality Management & Certifications

Quality System

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QMS (ISO, GMP)

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Quality Advantages

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Quality Control Process

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📂 Technical Resources & Downloads

PDF Product Manual ↓ PDF MSDS / SDS ↓

Frequently Asked Questions (FAQ)

Q1: What sample types can be tested with this kit?
This kit supports purified rAAV bulk, final product stocks and harvested cell-culture samples. For bulk or final product stocks, perform Benzonase treatment and nucleic acid extraction prior to qPCR. Cell culture samples may be extracted using the SHENTEK Virus DNA & RNA Extraction Kit.
Q2: What is the assay dynamic range and lowest quantifiable concentration?
The standard curve covers 2 × 10^1 to 2 × 10^6 copies/µL (20 to 2,000,000 copies/µL). The lowest standard used for quantitation (LOQ) is 20 copies/µL.
Q3: How is rcAAV contamination rate calculated?
rcAAV contamination rate = (Measured copies of Target gene) ÷ (Measured copies of Reference gene × dilution factor ÷ 2). Note that each rAAV contains two copies of the Reference gene.
Q4: Do I need to add ROX to every instrument?
No. Addition of 100× ROX is instrument-dependent. ABI 7500 requires 0.04 µL ROX per reaction for the Target assay; SHENTEK-96S, Roche 480 and some other systems do not require ROX. Follow your instrument documentation.

Research Use Only

✔

Research Use Only (RUO). For Research Use Only. Not for use in diagnostic procedures. – This product is intended for laboratory research purposes only and not for clinical or regulatory diagnostic use.

✔

Not intended for use in USDA or FDA regulated diagnostic testing or official compliance testing.

When can I expect my order to ship?

Most orders are filled and shipped within 2-3 business days from the time they are received.

Our standard shipping usually take 2-5 days.

We also provide express shippping for time-sensitive deliveries. 

Email contact@biofargo.com if you have any requirements.

 

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