Product Description

SHENTEK® Residual E. coli DNA Quantitation Kit is designed for quantitative measurement of residual E. coli host cell DNA in a variety of biopharmaceutical products. The kit uses quantitative PCR (qPCR) to enable rapid and specific quantitation at the femtogram level. It is suitable for a broad range of sample types from in-process samples to final products. The kit includes an E. coli Control DNA traceable to a primary DNA reference standard and a ready-to-use qPCR mix. For nucleic acid extraction, refer to the SHENTEK® Residual Host Cell DNA Sample Preparation Kit User Guide (Product No. SK030203D100).

Technical Specifications

Parameter	Details
Intended use	Quantitation of residual E. coli host cell DNA in biopharmaceutical samples (research use only).
Total reactions	Reagents for 100 reactions (as supplied with the kit).
Detection principle	Quantitative PCR (absolute quantitation using standard curve).
Dynamic range (standard curve)	Standard curve concentrations prepared: 3000, 300, 30, 3, 0.3, 0.03 pg/µL (recommended at least five concentration points).
Limit of quantitation / sensitivity	Reported sensitivity: femtogram level (kit statement). Practical lowest standard used: 0.03 pg/µL.
Reaction volume	qPCR reaction volume set to 30 µL (instrument setting described uses 30 µL).
qPCR cycling conditions	Activation: 95 °C 10:00 (1 cycle); Denaturation: 95 °C 00:15 (40 cycles); Annealing/extension: 60 °C 01:00 (read fluorescence during this step).
Analysis settings / threshold	Manual Ct with Threshold = 0.02; Automatic baseline recommended (ABI7500 SDS v1.4 example).
Negative and control acceptance criteria	ERC recovery rate acceptance: 50%–150%; NCS Ct should be larger than mean Ct of the lowest standard; NTC should be undetermined or Ct ≥ 35.00.
Storage stability	Kit components can be stored under appropriate conditions for up to 24 months (see individual component labels).
Compatible instruments	Compatible with common real-time PCR instruments including SHENTEK-96S, ABI 7500, Bio-Rad CFX96, LineGene 9600plus, Mx3000P, qTOWER3G, etc.

Features

High sensitivity: Enables quantitation at the femtogram level; lowest standard shown at 0.03 pg/µL.
Traceable control: Includes E. coli Control DNA traceable to a primary DNA reference standard for reliable standard curve construction.
Ready-to-use qPCR mix: Supplied E. coli qPCR MIX is provided in ready-to-use format to simplify reaction setup.
Broad sample compatibility: Validated conceptually for a variety of biopharmaceutical sample types from in-process to final products (recommended to perform method validation for specific matrices).
Instrument compatibility: Protocol and plate layouts provided for a range of common real-time PCR instruments.

Applications

Quantitation of residual E. coli host cell DNA in biopharmaceutical development and manufacturing
In-process monitoring of residual host DNA during purification workflows
Final product release testing (research use applications)
Method validation and assay development for host cell DNA quantitation

Kit Contents

E. coli DNA Control (NNA002)-20 °C; 50 µL × 1 tube

E. coli qPCR MIX (NNC002)-20 °C; 1 mL × 2 tubes (protect from light)

DNA Dilution Buffer (DDB) (NND001)-20 °C; 1.5 mL × 3 tubes

* Total: 100 reactions | Method: qPCR (absolute quantitation using a standard curve)

Attention

• For Research Use Only (RUO) — not for diagnostic procedures.

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

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

• Prevent contamination: use RNase/DNase-free consumables, low-retention filter tips, UV-decontaminate surfaces and pipettes, and use separate areas for pre- and post-amplification steps.

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

• Store kit components at -20 °C and protect light-sensitive reagents as indicated.

• Include at least five standard concentrations for a reliable standard curve and perform method validation for your sample matrix.

• Acceptable ERC recovery range: 50%–150%; verify NTC is undetermined or Ct ≥ 35 and NCS Ct is larger than mean Ct of lowest standard.

• Follow instrument-specific software and parameter settings for data analysis (example settings provided for ABI 7500 SDS v1.4).

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 intended use of this kit?

The kit is intended for research quantitation of residual E. coli host cell DNA in biopharmaceutical samples using qPCR (Research Use Only).

Q2: What are the standard curve concentrations supplied in the protocol?

The protocol describes preparing standards at 3000, 300, 30, 3, 0.3 and 0.03 pg/µL (use at least five concentrations for the standard curve).

Q3: What acceptance criteria should I use for controls?

ERC recovery should be between 50% and 150%; NTC should be undetermined or Ct ≥ 35; NCS Ct should be larger than the mean Ct of the lowest standard concentration.

Q4: How should I set up the qPCR cycling program?

Recommended example: 95 °C for 10:00 (activation, 1 cycle); 95 °C for 0:15 (denaturation) and 60 °C for 1:00 (annealing/extension with fluorescence read) for 40 cycles. Reaction volume example: 30 µL. Adjust program per instrument requirements.

Research Use Only

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

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

Residual E. coli DNA Quantitation Kit SHENTEK®