Residual Sf9 & AcNPV DNA Quantitation Kit SHENTEK®

For Research Use Only (RUO)

$2,575.00

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Sku: 1101101
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⚡ Detection Method
qPCR
🕒 Assay Time
Refer to manual
🎯 Storage Temperature
-20 °C (stable for 24 months)

Product Description

SHENTEK® Residual Sf9&AcNPV DNA Quantitation Kit is designed for quantitative measurement of residual Sf9 and AcNPV host cell DNA in biopharmaceutical materials at different processing stages (in-process to final products). The kit uses a multiplex quantitative PCR (qPCR) assay with internal positive control (IPC) to provide rapid, specific and reliable quantitation down to femtogram-level sensitivity. Supplied reagents include DNA control standards for Sf9 and AcNPV, primer/probe mixes, IPC mix, qPCR reaction buffer and DNA dilution buffer. The kit is compatible with common real-time PCR instruments (e.g., SHENTEK-96S, ABI 7500, Bio-Rad CFX96, FQD-96A). Refer to the SHENTEK® Residual Host Cell DNA Sample Preparation Kit (Product No. 1104191) for extraction guidance.

Technical Specifications

Parameter Details
Dynamic range (Sf9) 300 pg/µL to 0.003 pg/µL (standard curve points: 300, 30, 3, 0.3, 0.03, 0.003 pg/µL)
Dynamic range (AcNPV) 5.14×10^6 to 5.14×10^1 copies/µL (equivalent mass: 20 to 0.0002 pg/µL; standard curve points provided for both copies/µL and pg/µL)
Analytical sensitivity Sensitivity down to femtogram (fg) level implied by standard curve lowest points (Sf9: 0.003 pg/µL = 3 fg/µL)
Reaction format & volumes Reaction total volume 30 µL per well (20 µL qPCR MIX + 10 µL sample).
qPCR MIX composition (per reaction) qPCR Reaction Buffer 15.9 µL; Sf9&AcNPV Primer&Probe MIX 2.8 µL; IPC MIX 1.3 µL (total 20 µL qPCR MIX)
Thermal cycling conditions Activation: 95 °C for 10:00 (1 cycle); Denaturation: 95 °C for 0:15; Annealing/Extension: 60 °C for 1:00 (read fluorescence). 40 cycles.
Instrument channels / dyes Sf9-DNA: FAM; AcNPV-DNA: CY5; IPC: VIC; Passive reference dye: ROX (ABI 7500 example).
Standard curve / quantitation settings Manual Ct with threshold 0.02 for Sf9-DNA, AcNPV-DNA and IPC; use Absolute Quantitation (Standard Curve) analysis.
Controls and acceptance criteria IPC: Ct(sample) should be within ±1.0 Ct of Ct(NCS) — large deviation indicates inhibition. NTC: Ct ≥ 35.00 or undetermined. NCS Ct should be larger than mean Ct of lowest standard and show VIC amplification.
Number of reactions per kit Reagents for 100 reactions (as shipped).
Storage stability Kit components can be stored under appropriate conditions for up to 24 months (see component-specific storage: most components -20 °C; protect qPCR Reaction Buffer from light).
Threshold / baseline guidance Threshold set to 0.02; Automatic baseline recommended when using provided settings.

Features

  • Multiplex qPCR: Simultaneous detection and quantitation of Sf9-DNA, AcNPV-DNA and IPC in one reaction using distinct fluorescent channels (FAM, CY5, VIC).
  • High sensitivity: Assay standard curve includes femtogram-level concentrations (Sf9 down to 0.003 pg/µL) enabling detection at fg scale.
  • Internal Positive Control (IPC): IPC provided to monitor PCR inhibition and overall reaction performance for each sample.
  • Ready-to-use mixes: Pre-formulated primer/probe mix, IPC mix and reaction buffer simplify setup and reduce pipetting errors.
  • Instrument compatibility: Validated examples and guidance provided for multiple real-time PCR platforms (e.g., SHENTEK-96S, ABI 7500, CFX96, FQD-96A).
  • Standardized quantitation: Supplied Sf9 & AcNPV DNA controls with serial dilution scheme for generating absolute standard curves (copies/µL and pg/µL).

Applications

  • Quantitation of residual Sf9 host cell DNA in biopharmaceutical process and final drug substance
  • Quantitation of residual Autographa californica multiple nucleopolyhedrovirus (AcNPV) DNA in bioprocess samples
  • In-process quality control and release testing for products produced in Sf9/baculovirus systems
  • Method validation and assay development for host-cell DNA monitoring

Kit Contents

Sf9&AcNPV DNA Control (NNA051), lyophilized powder × 1 tube-20 °C
qPCR Reaction Buffer (NNB001), 850 µL × 2 tubes-20 °C; protect from light
Sf9&AcNPV Primer&Probe MIX (NNC059), 300 µL × 1 tube-20 °C
IPC MIX (NNC067), 150 µL × 1 tube-20 °C
DNA Dilution Buffer (DDB) (NND001), 1.5 mL × 3 tubes-20 °C (unused remaining DDB may be stored at 2–8 °C; if cloudy, heat at 37 °C until clear)
* Total: 100 reactions | Method: qPCR (multiplex quantitative PCR, absolute quantitation via standard curve)

Attention

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

• Read Material Safety Data Sheets (MSDS) and follow handling instructions.

• Wear appropriate PPE (eyewear, mask, gloves, protective clothing).

• Work in DNase-free area; use DNase-free, low-retention consumables and tips.

• UV-decontaminate work surfaces and equipment for 30 minutes and disinfect with 75% ethanol before setup.

• Thaw reagents at 2–8 °C or on ice; vortex and spin briefly before use.

• Prepare standards by serial 10-fold dilutions in DDB as instructed; at least five standard concentrations recommended.

• Seal plates well, mix and spin down prior to starting qPCR to avoid evaporation and inconsistent Ct values.

• Monitor IPC: sample Ct-IPC should be within ±1.0 Ct of NCS; significant increase indicates sample inhibition.

• NTC should have Ct ≥ 35.00 or be undetermined; NCS Ct should be larger than mean Ct of lowest standard and show IPC amplification.

• Protect qPCR Reaction Buffer from light.

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: How many reactions does one kit support?
This kit provides reagents for 100 reactions (stated on the product cover).
Q2: What is the recommended reaction setup and thermal program?
Prepare 30 µL reactions (20 µL qPCR MIX + 10 µL sample). Thermal cycling: Activation 95 °C 10:00; 40 cycles of 95 °C 0:15 and 60 °C 1:00 with fluorescence read during the 60 °C step. Use manual Ct threshold 0.02 as guidance.
Q3: What should I do if IPC indicates inhibition?
If sample Ct-IPC is significantly higher than Ct-IPC of NCS (beyond ±1.0 Ct), consider sample dilution, re-extraction, or purification to remove inhibitors. Use recovery of spiked reference standards to assess sample processing.
Q4: What are the assay dynamic ranges and units?
Sf9 standard curve: 300 to 0.003 pg/µL. AcNPV standard curve: 5.14×10^6 to 5.14×10^1 copies/µL (equivalent to 20 to 0.0002 pg/µL). At least five standard concentrations should be used for curve generation.

Research Use Only

✔

Research Use Only (RUO) – 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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