Residual SV40 LTA/E1A DNA Quantitation Kit (2G) SHENTEK™

For Research Use Only (RUO)

$2,575.00

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Sku: 1403443
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⚡ Detection Method
qPCR
🕒 Assay Time
Refer to manual
🎯 Storage Temperature
–20 °C (stable for 24 months)

Product Description

SHENTEK® Residual SV40LTA & E1A DNA Quantitation Kit (2G) is a duplex real-time PCR (qPCR) kit designed to quantify residual SV40 large T antigen (LTA) and E1A host cell DNA (e.g., from HEK293T). The assay is duplex (simultaneous detection of SV40LTA and E1A) with an internal positive control (IPC) and achieves sensitivity to the level of 10^1 copies/μL. The kit includes linear and non-linear DNA controls for standard curve generation and is compatible with common real-time PCR platforms (SHENTEK-96S, ABI 7500, Bio-Rad CFX96). Reagents are provided for 100 reactions.

Technical Specifications

Parameter Details
Intended use Quantitation of residual SV40LTA and E1A host cell DNA in biological samples
Detection principle Duplex real-time PCR (qPCR) with target-specific primer/probe mixes and IPC
Sensitivity (LOD) 10^1 copies/μL (stated detection level)
Standard curve dynamic range (linear DNA) Typical standard curve points: 4.67 × 10^6 to 4.67 × 10^1 copies/μL (ST1–ST6)
Standard curve dynamic range (non-linear DNA) Typical standard curve points: 2.80 × 10^6 to 2.80 × 10^2 copies/μL (ST1–ST5)
Control concentrations (as supplied) Linear DNA Control: SV40LTA 4.67 × 10^9 copies/μL; E1A 4.97 × 10^9 copies/μL. Non-linear DNA Control: SV40LTA 2.80 × 10^9 copies/μL; E1A 2.98 × 10^9 copies/μL
Quantitation format Absolute quantitation using standard curve (user-prepared serial dilutions)
Reaction setup / volume Total reaction volume 30 μL (20 μL qPCR MIX + 10 μL sample or standard). qPCR MIX per reaction: qPCR Reaction Buffer 15.9 μL, Primer & Probe MIX 2.8 μL, IPC MIX 1.3 μL (total 20 μL)
Thermal cycling Activation 95 °C 10:00 (1 cycle); Denaturation 95 °C 0:15; Annealing/extension 60 °C 1:00; 40 cycles. Fluorescence read during annealing/extension
Instrument compatibility SHENTEK-96S, ABI 7500 (SDS v1.4 example), Bio-Rad CFX96 (other real-time PCR instruments supported with appropriate settings)
Analysis settings (recommended) Manual Ct with threshold = 0.02 and Automatic Baseline; set Standard Curve (Absolute Quantitation) analysis
IPC acceptance criteria Mean Ct(IPC) of sample should be within ±1.0 of the NCS Ct(IPC); significant increase indicates potential inhibition
NTC acceptance thresholds SV40LTA detection value of NTC should be ≤ 14.32 copies/μL; E1A detection value of NTC should be ≤ 17.79 copies/μL (or use user-validated criteria)
Provided reactions Reagents for 100 reactions
Shelf life Kit components can be stored under recommended conditions for up to 24 months

Features

  • Duplex real-time PCR: Simultaneous quantitation of SV40LTA and E1A targets in the same reaction using target-specific probes and an IPC.
  • High sensitivity: Detects down to 10^1 copies/μL enabling detection of low-level residual host cell DNA.
  • Ready-to-use controls and mixes: Includes linear and non-linear DNA controls, qPCR reaction buffer, primer/probe mix, and IPC mix to support standard curve generation and assay QC.
  • Platform compatibility: Validated on common qPCR instruments (SHENTEK-96S, ABI 7500, Bio-Rad CFX96) and adaptable to other real-time PCR systems.
  • Pre-defined analysis settings: Recommended thermal cycling and analysis parameters (e.g., threshold = 0.02, manual Ct) provided to standardize quantitation.

Applications

  • Quantitation of residual SV40LTA and E1A host cell DNA in cell substrates (e.g., HEK293T)
  • Residual host cell DNA monitoring for upstream/downstream bioprocessing and product release testing (research use)
  • Method validation and standard curve generation for absolute quantitation of SV40-derived sequences

Kit Contents

SV40LTA & E1A linear DNA Control (NNA019) — lyophilized powder ×1 tube-20 °C
SV40LTA & E1A non-linear DNA Control (NNA020) — 50 μL ×1 tube-20 °C
qPCR Reaction Buffer (NNB001) — 850 μL ×2 tubes-20 °C, protect from light
SV40LTA & E1A Primer & Probe MIX (NNC030) — 300 μL ×1 tube-20 °C, protect from light
IPC MIX (NNC066) — 150 μL ×1 tube-20 °C, protect from light
DNA Dilution Buffer (DDB) (NND001) — 1.5 mL ×3 tubes-20 °C (store remaining DDB at 2–8 °C after opening per protocol)
* Total: 100 reactions | Method: qPCR

Attention

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

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

• Wear appropriate PPE: protective eyewear, mask, laboratory coat, and gloves.

• Avoid contamination: irradiate bench, pipettes and tubes with UV for 30 minutes and disinfect surfaces with 75% ethanol before setup.

• Reconstitute lyophilized control carefully: add 55 μL ddH2O, mix and let stand 10 minutes; perform serial dilutions as instructed.

• Protect light-sensitive reagents (qPCR Reaction Buffer, Primer & Probe MIX, IPC MIX) from light and store at –20 °C.

• Prepare at least five concentration points for the standard curve; perform method validation to select appropriate sample dilutions.

• DDB: store remaining unused DDB at 2–8 °C; if cloudy, heat to 37 °C until clear.

• IPC criteria: mean Ct(IPC) should be within ±1.0 of NCS Ct(IPC); significant shifts indicate inhibition.

• NTC acceptance: SV40LTA ≤ 14.32 copies/μL and E1A ≤ 17.79 copies/μL (or use user-defined validation criteria).

• Configure analysis parameters per instrument and software version; recommended settings include manual Ct and threshold = 0.02.

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 limit of detection (LOD) for this kit?
The kit reports sensitivity to the level of 10^1 copies/μL. Actual LOD/LOQ should be confirmed during your laboratory validation.
Q2: Which instruments are compatible with this kit?
The kit is compatible with common real-time PCR instruments including SHENTEK-96S, ABI 7500 (SDS v1.4 example) and Bio-Rad CFX96. Other instruments may be used with appropriate program and analysis adjustments.
Q3: How should I set up the qPCR thermal profile and analysis?
Recommended program: Activation 95 °C 10:00; 40 cycles of 95 °C 0:15 and 60 °C 1:00 (read fluorescence during annealing/extension). Analysis: Manual Ct with threshold = 0.02 and Automatic Baseline; use Absolute Quantitation (standard curve).
Q4: What controls should I include and what are acceptance criteria?
Include NTC, NCS, and at least five standard curve points (ST1–ST5; ST6 optional for linear control). IPC mean Ct should be within ±1.0 of NCS. NTC detection values should be ≤ 14.32 copies/μL for SV40LTA and ≤ 17.79 copies/μL for E1A, or use your validated criteria.

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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