Residual Hi5 & AcNPV DNA Quantitation Kit SHENTEK®

For Research Use Only (RUO)

$2,575.00

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Sku: 1101102
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⚡ Detection Method
qPCR
🕒 Assay Time
Refer to manual
🎯 Storage Temperature
–20 °C (stable for 24 months)

Product Description

SHENTEK® Residual Hi5&AcNPV DNA Quantitation Kit is designed to quantify residual Hi5 and AcNPV host-cell DNA in biopharmaceutical samples (in-process and final products). The kit uses a multiplex quantitative PCR (qPCR) assay with target-specific primer/probe mixes for Hi5 DNA and AcNPV DNA plus an Internal Positive Control (IPC) to monitor reaction performance and potential inhibition. The assay supports absolute quantitation using a supplied lyophilized DNA control and DDB-based serial dilutions. The kit provides reagents for 100 reactions and is compatible with common real-time PCR instruments (e.g., SHENTEK-96S, ABI 7500, Bio-Rad CFX96, FQD-96A).

Technical Specifications

Parameter Details
Intended use Quantitation of residual Hi5 and AcNPV host-cell DNA in biopharmaceutical samples across process stages
Assay format Multiplex quantitative PCR (qPCR) with three detectors: Hi5-DNA (FAM), AcNPV-DNA (CY5), IPC (VIC); ROX as passive reference
Reaction volume 30 μL total per well (20 μL qPCR mix + 10 μL sample)
qPCR mix composition (per reaction) qPCR Reaction Buffer 15.9 μL; Hi5&AcNPV Primer&Probe MIX 2.8 μL; IPC MIX 1.3 μL (total 20 μL)
Thermal cycling conditions Activation 95 °C 10:00; 40 cycles of 95 °C 00:15 and 60 °C 01:00 (fluorescence read at 60 °C)
Hi5 DNA standard curve (mass) Standard points: 30 pg/μL (ST1), 3 pg/μL (ST2), 0.3 pg/μL (ST3), 0.03 pg/μL (ST4), 0.003 pg/μL (ST5)
AcNPV DNA standard curve Standard points (copies/μL and pg/μL): ST1 5.14×10^5 copies/μL (2 pg/μL), ST2 5.14×10^4 (0.2 pg/μL), ST3 5.14×10^3 (0.02 pg/μL), ST4 5.14×10^2 (0.002 pg/μL), ST5 5.14×10^1 (0.0002 pg/μL)
Sensitivity claim Quantitation at femtogram (fg) level is supported by the provided standard curve down to 0.003 pg/μL for Hi5 and 0.0002 pg/μL (5.14×10^1 copies/μL) for AcNPV
Controls and acceptance criteria IPC Ct of sample vs NCS should be within ±1.0 Ct; NTC Ct should be ≥35 or undetermined; NCS Ct should be larger (i.e., later) than mean Ct of lowest standard; NCS should show VIC amplification
Sample input volume 10 μL purified test sample (per reaction)
Kit stability Components stable up to 24 months when stored as directed
Provided quantity Reagents sufficient for 100 reactions (kit contains reagents sized as listed in kit contents)
LOD / LOQ Validated LOQ is referenced in method validation; the user should determine LOQ during method validation. If LOQ < lowest standard, NCS value should be less than LOQ

Features

  • Multiplex qPCR format: Simultaneous detection of Hi5 DNA, AcNPV DNA and IPC in a single reaction (FAM, CY5, VIC channels)
  • High sensitivity: Supports quantitation down to femtogram-level concentrations using supplied standards (mass and copy-number options for AcNPV)
  • Internal positive control (IPC): IPC included to monitor PCR performance and identify inhibited samples
  • Ready-to-use mixes: Pre-formulated qPCR Reaction Buffer, Primer&Probe MIX and IPC MIX reduce assay setup time and variability
  • Compatible with common qPCR instruments: Validated on multiple real-time PCR platforms (SHENTEK-96S, ABI 7500, Bio-Rad CFX96, FQD-96A)

Applications

  • Quantitation of residual Hi5 host-cell DNA in biopharmaceutical process development and quality control
  • Quantitation of residual AcNPV DNA (baculovirus) in upstream and downstream process samples
  • In-process monitoring of host DNA clearance during purification workflows
  • Release testing support and method development for biologics produced in Hi5/AcNPV systems

Kit Contents

Hi5&AcNPV DNA Control (lyophilized powder) ×1 tube–20 °C
Hi5&AcNPV Primer&Probe MIX 300 μL ×1 tube–20 °C, protect from light
qPCR Reaction Buffer 850 μL ×2 tubes–20 °C
IPC MIX 150 μL ×1 tube–20 °C
DNA Dilution Buffer (DDB) 1.5 mL ×3 tubes–20 °C (after opening/for use, store remaining DDB at 2–8 °C)
* Total: 100 reactions | Method: qPCR

Attention

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

• Read Material Safety Data Sheets (MSDS) and wear appropriate PPE (gloves, mask, protective eyewear, lab coat).

• Prevent nucleic acid contamination: irradiate work surfaces, micropipettes and tubes with UV for 30 minutes and disinfect with 75% ethanol prior to setup.

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

• Follow plate layout recommendations and include at least five standard curve points, NTC and NCS in each run.

• IPC acceptance: Ct(sample IPC) must be within ±1.0 Ct of Ct(NCS) — higher IPC Ct indicates potential inhibition.

• NTC acceptance: Ct ≥ 35 or undetermined; NTC should not show target amplification.

• If DNA Dilution Buffer (DDB) is cloudy, heat at 37 °C until clear before use.

• Validated LOQ must be established during method validation; if LOQ is below the lowest standard, ensure NCS value is below LOQ.

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 is the minimum detectable concentration for Hi5 and AcNPV using this kit?
The provided standard curve supports quantitation down to 0.003 pg/μL for Hi5 DNA and 0.0002 pg/μL (5.14×10^1 copies/μL) for AcNPV DNA. Users should perform method validation to determine assay LOD/LOQ for their matrix.
Q2: How much sample should be added per reaction?
Add 10 μL of purified test sample per 30 μL reaction (20 μL qPCR mix + 10 μL sample).
Q3: What are the instrument settings for the assay?
Thermal profile: Activation 95 °C for 10:00; 40 cycles of 95 °C for 00:15 and 60 °C for 01:00 with fluorescence read at 60 °C. Set reaction volume to 30 μL. Configure detectors: Hi5-DNA (FAM), AcNPV-DNA (CY5), IPC (VIC); ROX as passive reference.
Q4: What should I do if IPC indicates inhibition?
If the sample IPC Ct is significantly higher than NCS IPC Ct (outside ±1.0 Ct), suspect inhibition. Consider diluting the sample, re-extracting with improved purification, or including spike-recovery controls to assess sample recovery.

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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