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Product Description
SHENTEK® Residual HEK293 DNA Size Analysis Kit is designed to quantify residual HEK293 host cell DNA fragments of different sizes across biopharmaceutical sample types (in-process samples to final products) using real-time PCR (qPCR). The kit amplifies four fragment sizes (75 bp, 122 bp, 244 bp and 562 bp) to determine size distribution and quantity down to the femtogram (fg) level. The kit includes primer/probe mixes for each fragment, internal positive control (IPC), DNA dilution buffer and a HEK293 DNA control for standard curve preparation. For extraction guidance, refer to the SHENTEK® Residual Host Cell DNA Sample Preparation Kit (Product No. 1104191).
Technical Specifications
| Parameter | Details |
|---|---|
| Target fragment sizes | 75 bp, 122 bp, 244 bp, 562 bp |
| Analytical method | Real-time PCR (qPCR) with TaqMan-style primer & probe mixes and IPC |
| Quantitation sensitivity | Designed for quantitation at the femtogram (fg) level (lowest standard in kit workflow corresponds to 0.03 pg/µL = 30 fg) |
| Standard curve concentrations (prepared from supplied HEK293 DNA Control) | ST1 = 300 pg/µL; ST2 = 30 pg/µL; ST3 = 3 pg/µL; ST4 = 0.3 pg/µL; ST5 = 0.03 pg/µL (ST0 = 3000 pg/µL stock prior to dilution) |
| Linear / working range (as provided by recommended standards) | Demonstrated standard points from 0.03 pg/µL to 300 pg/µL (prepare standard curve using ST1–ST5); ST0 (3000 pg/µL) used as stock |
| qPCR reaction volume | 30 μL total reaction volume (20 μL qPCR master mix + 10 μL template) |
| qPCR master mix composition (per reaction) | qPCR Reaction Buffer 15.9 μL; HEK293 Primer&Probe MIX (appropriate fragment) 2.8 μL; IPC MIX 1.3 μL; total 20 μL (then add 10 μL template) |
| Thermal cycling conditions | Activation: 95 °C 10:00 (1 cycle); Denaturation: 95 °C 00:15; Annealing: 60 °C 00:30; Extension/read: 72 °C 01:30; 40 cycles |
| Instrument compatibility | SHENTEK-96S, Applied Biosystems 7500, Bio-Rad CFX96, FQD-96A and similar real-time PCR instruments (user must configure detectors/channels accordingly) |
| IPC / quality control criteria | Sample Ct-IPC should be within ±1.0 of the NCS Ct-IPC. Ct of NTC and NCS should be larger than the mean Ct of the lowest concentration standard. |
| Kit stability / shelf life | Kit components can be stored under appropriate conditions for up to 24 months (check label expiration date) |
Features
- Size-resolved quantitation: Simultaneous assays for four discrete fragment lengths (75, 122, 244, 562 bp) allow determination of residual HEK293 DNA size distribution.
- High sensitivity: Designed to quantify DNA down to the femtogram range (lowest recommended standard 0.03 pg/µL = 30 fg).
- Ready-to-use primer & probe mixes: Pre-formulated HEK293 Primer&Probe MIXes for each fragment size reduce setup time and minimize variability.
- Internal process control (IPC): IPC MIX included to monitor inhibition and assay performance; acceptance criteria provided.
- Flexible instrument compatibility: Compatible with multiple real-time PCR platforms (instructions include detector/channel setup and reference dye settings).
- Complete workflow support: Includes DNA dilution buffer and HEK293 DNA control for standard curve preparation; refers to complementary sample prep kit for extraction.
Applications
- Quantitation of residual HEK293 host cell DNA in biopharmaceutical process and final product testing
- Determination of host cell DNA fragment size distribution
- In-process monitoring and process development for biologics produced in HEK293 cells
- Release testing and quality control (research-use contexts)
Kit Contents
qPCR Reaction Buffer (NNB001), 850 μL × 8 tubes-20 °C, protect from light
HEK293 Primer&Probe MIX-75 (NNC012), 300 μL × 1 tube-20 °C
HEK293 Primer&Probe MIX-122 (NNC013), 300 μL × 1 tube-20 °C
HEK293 Primer&Probe MIX-244 (NNC014), 300 μL × 1 tube-20 °C
HEK293 Primer&Probe MIX-562 (NNC015), 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 (store remaining DDB at 2–8 °C after thaw; if cloudy, heat at 37 °C until clear)
HEK293 DNA Control (NNA005), 50 μL × 1 tube-20 °C
* Total: 4 × 100 reactions | Method: qPCR
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, lab coat).
• Prevent nucleic acid contamination: irradiate work surfaces, pipettes and tubes with UV for 30 minutes and disinfect with 75% ethanol.
• Thaw reagents at 2–8 °C or on ice; vortex and briefly spin down before use.
• Prepare separate standard curves for each fragment-specific primer/probe mix (75, 122, 244, 562 bp).
• Include at least five standard points (recommended ST1–ST5) and run samples in triplicate where possible.
• qPCR run setup: reaction volume 30 μL (20 μL master mix + 10 μL template); thermal cycling and detector settings must be configured per instrument.
• IPC acceptance: sample Ct-IPC should be within ±1.0 of NCS Ct-IPC — higher Ct-IPC indicates potential inhibition; consider recovery controls.
• Negative controls (NTC) and NCS should show Ct values greater (later) than the mean Ct of the lowest standard point.
Quality Management & Certifications
Quality System
Download QMSQMS (ISO, GMP)
Download CertificateQuality Advantages
View ReportQuality Control Process
Download ProcessTechnical Resources & Downloads
Frequently Asked Questions (FAQ)
Q1: What is the sensitivity / limit of detection (LOD) of this kit?
The kit is designed to quantify HEK293 DNA at the femtogram level. The supplied standard curve includes points down to 0.03 pg/µL (30 fg); actual LOD/LOQ should be established during method validation in your laboratory.
Q2: Which fragment sizes are measured and how are they assayed?
Four fragment sizes are assayed separately using fragment-specific primer & probe mixes: 75 bp, 122 bp, 244 bp and 562 bp. Prepare four separate standard curves (one per fragment) and run each assay on the qPCR instrument.
Q3: Which instruments are compatible?
The kit is compatible with common real-time PCR systems such as SHENTEK-96S, Applied Biosystems 7500, Bio-Rad CFX96, and FQD-96A. Users must configure detectors/channels, passive reference dye (e.g., ROX if required), and cycling conditions according to their instrument.
Q4: How do I know if a sample is inhibited?
Check the IPC (internal positive control). The sample Ct-IPC should be within ±1.0 of the NCS Ct-IPC. A significantly higher Ct-IPC in the sample suggests inhibition; dilute the sample or perform recovery controls and re-test.
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.
