Residual E. coli DNA Size Analysis Kit (2G) SHENTEK®

For Research Use Only (RUO)

$3,708.00

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Sku: 1103171-2
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⚡ Detection Method
qPCR
🕒 Assay Time
Refer to manual
🎯 Storage Temperature
-20 °C (stable for 24 months)

Product Description

SHENTEK® Residual E.coli DNA Size Analysis Kit (2G) is a real-time PCR (qPCR) kit for rapid and specific quantitation of residual Escherichia coli (E.coli) host-cell DNA across different fragment sizes during biopharmaceutical development and manufacturing. The kit amplifies four target fragment lengths (85 bp, 103 bp, 220 bp and 550 bp) using FAM-labeled probes and includes an internal positive control (IPC, VIC) for inhibition monitoring. It supports absolute quantitation via user-prepared standard curves prepared from the supplied E.coli DNA Control and DNA Dilution Buffer (DDB). The kit is supplied as ready-to-use primer/probe mixes and reaction buffer for high-throughput testing and is compatible with common real-time PCR instruments (e.g., SHENTEK-96S, ABI 7500, Bio-Rad CFX96, LineGene 9600). For extraction/preparation of host-cell DNA prior to qPCR, refer to the SHENTEK® Residual Host Cell DNA Sample Preparation Kit (Product No. 1104191).

Technical Specifications

Parameter Details
Target fragment sizes 85 bp, 103 bp, 220 bp, 550 bp
Quantitation range (standards provided) Serial standard curve points prepared from E.coli DNA Control: ST1 = 300 pg/µL, ST2 = 30 pg/µL, ST3 = 3 pg/µL, ST4 = 0.3 pg/µL, ST5 = 0.03 pg/µL (prepare from ST0 = 3000 pg/µL)
Implied sensitivity / lowest standard Lowest standard = 0.03 pg/µL (30 fg/µL); kit described as detecting at the femtogram (fg) level
Reaction volumes qPCR MIX per well: 20 µL; add 10 µL sample/standard/NTC/NCS for a total reaction volume of 30 µL
qPCR thermal cycling Activation: 95 °C 10:00 (1 cycle); Denaturation: 95 °C 00:15; Annealing: 60 °C 00:30; Extension: 72 °C 01:30; 40 cycles. Instrument reads fluorescence during extension step.
Fluorescent channels / detectors Targets (E.coli fragments) - FAM; IPC - VIC; Passive reference - ROX
Standard curve / analysis settings (example) Use Absolute Quantitation (Standard Curve). Analysis settings example: Manual Ct; Threshold = 0.02; Automatic baseline. Verify slope, intercept and R2 values.
IPC acceptance criteria Mean Ct of IPC for sample should be within ±1.0 Ct of NCS Ct-IPC. Significant increase indicates possible inhibition.
NTC / NCS criteria Ct of NTC and NCS should be larger than mean Ct of lowest standard or show no significant amplification in FAM channel while showing normal VIC signal.
Shelf life Kit components: up to 24 months (check label expiration date)
Compatible instruments SHENTEK-96S, ABI 7500, Bio-Rad CFX96, LineGene 9600 and other real-time PCR systems

Features

  • Multiplex fragment-size analysis (four assays): Designed to quantify four discrete E.coli DNA fragment lengths (85, 103, 220 and 550 bp) to determine size distribution of residual host DNA.
  • High sensitivity: Detection capability down to femtogram (fg) level; lowest provided standard 0.03 pg/µL (30 fg/µL).
  • Ready-to-use primer/probe mixes: Pre-formulated Primer & Probe MIX for each fragment (85, 103, 220, 550) simplifies setup and reduces handling error.
  • Internal positive control (IPC): IPC (VIC) included to monitor inhibition and sample integrity; IPC acceptance criteria provided.
  • Compatibility and throughput: Compatible with multiple real-time PCR platforms and formatted for 96-well plates; reagents sufficient for 4 × 100 reactions (high-throughput).
  • Absolute quantitation: Supplied E.coli DNA Control and dilution scheme enables generation of standard curves for absolute quantitation.

Applications

  • Quantitation of residual E.coli host-cell DNA in biopharmaceutical process and final products
  • In-process monitoring of host DNA clearance during process development
  • Quality control and characterization of DNA fragment size distribution in purified biologics
  • Method validation and assay development for host-cell DNA assessment (research use)

Kit Contents

E.coli DNA Control (NNA002) 50 μL × 1 tube-20 °C
qPCR Reaction Buffer (NNB001) 850 μL × 8 tubes-20 °C (protect from light)
E.coli Primer & Probe MIX-85 (NNC110) 300 μL × 1 tube-20 °C
E.coli Primer & Probe MIX-103 (NNC111) 300 μL × 1 tube-20 °C
E.coli Primer & Probe MIX-220 (NNC112) 300 μL × 1 tube-20 °C
E.coli Primer & Probe MIX-550 (NNC113) 300 μL × 1 tube-20 °C
IPC MIX (NNC069) 550 μL × 1 tube-20 °C
DNA Dilution Buffer (DDB) (NND001) 1.5 mL × 3 tubes-20 °C (after thaw, store at 2–8 °C; if cloudy heat to 37 °C until clear)
* Total: 400 reactions (4 × 100) | Method: qPCR

Attention

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

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

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

• Prevent contamination: use DNase-free, low-retention consumables; irradiate work surfaces and pipettes with UV for 30 minutes and disinfect with 75% ethanol prior to setup.

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

• Prepare separate standard curves for each fragment length (four assays).

• qPCR reaction setup: prepare qPCR MIX (20 µL) and add 10 µL sample/standard for 30 µL total reaction volume.

• Analysis guidance: use Manual Ct, Threshold = 0.02 (example); verify slope, intercept and R2 of standard curve.

• IPC acceptance: mean Ct-IPC for sample should be within ±1.0 Ct of NCS Ct-IPC; elevated IPC Ct suggests inhibition.

• NTC/NCS should show no significant amplification in FAM channel or Ct greater than lowest standard; VIC signal should be present for IPC.

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 use to quantify residual E.coli host-cell DNA and determine fragment size distribution in biopharmaceutical samples (in-process and final products). It is RUO and not for clinical diagnostic use.
Q2: What instrument channels should I use for the assay?
Use FAM channel(s) for E.coli target fragment detection, VIC for the internal positive control (IPC), and ROX as the passive reference according to your instrument configuration.
Q3: What is the lowest concentration the kit can detect?
The supplied standard curve includes a lowest standard of 0.03 pg/µL (30 fg/µL). The kit description indicates detection at the femtogram level; actual limit of detection should be confirmed during in-house validation.
Q4: How should I prepare the standards and controls?
Thaw E.coli DNA Control and DDB at 2–8 °C or on ice. Dilute DNA Control to ST0 = 3000 pg/µL then prepare serial dilutions to produce ST1–ST5 (300, 30, 3, 0.3, 0.03 pg/µL). Include at least five concentrations, 1 NTC and 1 NCS per run; run samples in triplicate when possible.

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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