0 item(s)
View cart
You have no items in your shopping cart.
Product Description
SHENTEK® Residual BHK DNA Size Analysis Kit is designed for quantitative measurement of residual BHK host-cell DNA fragments across different fragment sizes in biopharmaceutical samples (from in-process samples to final products). The assay uses quantitative PCR with FAM-labeled probes to amplify four discrete fragment sizes (81 bp, 134 bp, 216 bp and 589 bp) to determine size distribution and quantity. The kit provides high sensitivity with a reported limit of detection (LOD) at the femtogram (fg) level. An internal positive control (IPC, VIC channel) is included to monitor inhibition and extraction/reaction performance. For DNA extraction, refer to the SHENTEK® Residual Host Cell DNA Sample Preparation Kit (Product No. 1104191).
Technical Specifications
| Parameter | Details |
|---|---|
| Target fragment sizes | 81 bp, 134 bp, 216 bp, 589 bp |
| Limit of detection (LOD) | Femtogram (fg) level sensitivity (LOD stated as fg level) |
| Standard curve concentrations (per kit instructions) | Standards used (ST1-ST5): 300 pg/µL, 30 pg/µL, 3 pg/µL, 0.3 pg/µL, 0.03 pg/µL (serial dilutions prepared from ST0 source) |
| Reaction volumes | qPCR reaction volume: 30 µL total (20 µL qPCR MIX + 10 µL sample/standard) |
| qPCR mix composition (per reaction) | qPCR Reaction Buffer 15.9 µL; BHK Primer & Probe MIX (per fragment) 2.8 µL; IPC MIX 1.3 µL; total 20 µL qPCR MIX |
| Thermal cycling conditions | Activation: 95 °C 10:00 (1 cycle); Denaturation: 95 °C 00:15; Annealing: 60 °C 00:30; Extension: 72 °C 01:30; 40 cycles. Fluorescence is read during the extension step. |
| Detection channels / probes | Target probes: FAM reporter for each BHK fragment assay; IPC: VIC reporter. ROX recommended as passive reference (ABI instruments). |
| NTC / NCS acceptance criteria | NTC Ct should be ≥ 35 cycles; NCS Ct should be greater than mean Ct of the lowest standard and show normal VIC amplification. IPC Ct of sample should be within ±1.0 of NCS IPC Ct; deviations indicate inhibition. |
| Recovery acceptance | Sample recovery rate (ERC-based) should be between 50% and 150% |
| Kit stability | Kit components can be stored under appropriate conditions for up to 24 months (see component labels) |
| Assay throughput | Reagents provided for 4 × 100 reactions (stated in product label / packaging) |
| LOQ | Limit of quantitation (LOQ) not numerically specified in the user guide; users should refer to assay validation for LOQ |
Features
- Multi‑size quantitation: Simultaneous assessment of residual BHK DNA across four discrete fragment sizes (81, 134, 216, 589 bp) to provide size distribution information.
- High sensitivity: Assay sensitivity reaches femtogram (fg) level LOD for detection of low-abundance residual DNA.
- qPCR-based specificity: TaqMan-style primer/probe assays (FAM) deliver rapid and specific quantitation; IPC (VIC) monitors inhibition and reaction integrity.
- Ready-to-use mixes: Pre-formulated qPCR Reaction Buffer and Primer&Probe MIXes simplify assay setup and reproducibility.
- Instrument compatibility: Validated for common real-time PCR platforms (examples: SHENTEK-96S, ABI 7500, Roche LightCycler 480) and configurable for other real-time PCR systems.
Applications
- Quantitation of residual BHK host-cell DNA in biopharmaceutical process samples
- Size-distribution analysis of residual DNA in in-process and final drug product samples
- Method development and validation for host-cell DNA monitoring
- Quality control testing during upstream/downstream processing and release testing (research use)
Kit Contents
BHK DNA Control (NNA029) 50 µL × 1 tube-20 °C
qPCR Reaction Buffer (NNB001) 850 µL × 8 tubes-20 °C (protect from light)
BHK Primer & Probe MIX-81 (NNC072) 300 µL × 1 tube-20 °C
BHK Primer & Probe MIX-134 (NNC073) 300 µL × 1 tube-20 °C
BHK Primer & Probe MIX-216 (NNC074) 300 µL × 1 tube-20 °C
BHK Primer & Probe MIX-589 (NNC075) 300 µL × 1 tube-20 °C
IPC MIX (NNC070) 550 µL × 1 tube-20 °C
DNA Dilution Buffer (DDB) (NND001) 1.5 mL × 3 tubes-20 °C
* Total: Reagents for 4 × 100 reactions (400 total reactions, as stated on product label) | Method: qPCR
Attention
• For research use only (see compliance). Not for diagnostic or therapeutic use.
• Read Material Safety Data Sheets (MSDS) and follow handling instructions. Wear appropriate PPE (eyewear, mask, gloves, protective clothing).
• Avoid nucleic acid contamination: irradiate work surfaces, pipettes and tubes with UV for 30 minutes and disinfect with 75% ethanol before setup.
• Thaw reagents at 2–8 °C (on ice), vortex and briefly centrifuge before use; protect qPCR Reaction Buffer from light.
• Prepare standard curves (minimum five concentrations recommended) and run samples in triplicate when possible.
• qPCR reaction setup: 20 µL qPCR MIX + 10 µL sample/standard = 30 µL total per well; seal plates and centrifuge briefly prior to run.
• Acceptance criteria: NTC Ct ≥ 35; IPC Ct of sample should be within ±1.0 of NCS IPC Ct; recovery rates should be between 50% and 150%.
• If DNA Dilution Buffer (DDB) is cloudy, heat at 37 °C until clear and store remaining DDB at 2–8 °C.
• Limit of quantitation (LOQ) must be established/validated by user; if LOQ is lower than lowest standard, NCS Ct should be below LOQ concentration.
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 fragment sizes does this kit detect and quantify?
The kit amplifies and quantifies four discrete BHK host‑cell DNA fragment sizes: 81 bp, 134 bp, 216 bp and 589 bp.
Q2: What is the assay sensitivity (LOD) of the kit?
The user guide reports a limit of detection (LOD) at the femtogram (fg) level. Exact LOD/LOQ values should be confirmed during in-house validation.
Q3: What reaction volume and thermal cycling program should I use?
Use a 30 µL reaction (20 µL qPCR MIX + 10 µL sample). Thermal cycling: Activation 95 °C 10:00; 40 cycles of 95 °C 00:15, 60 °C 00:30, 72 °C 01:30. Fluorescence is read during the extension step.
Q4: How should I interpret IPC, NTC and NCS results?
NTC Ct should be ≥ 35 cycles. IPC Ct of a sample should be within ±1.0 of the NCS IPC Ct; a higher IPC Ct indicates potential inhibition. NCS Ct should be greater than the mean Ct of the lowest standard and exhibit normal VIC amplification. Recovery rate (ERC-based) should be 50–150%.
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.
