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For Research Use Only (RUO)
$1,010.00
Sku: SK030203D100
Product Description
SHENTEK® Residual Host Cell DNA Sample Preparation Kit is a magnetic-particle based sample preparation kit designed for efficient and reproducible recovery of trace residual host-cell DNA from a wide range of biological products and complex sample matrices across multiple manufacturing steps. The kit is compatible with manual workflows or automated extraction using the rHCDpurify instrument and is intended to be used with SHENTEK® host cell DNA quantitation and size analysis kits. The kit supports multiple host cell types (CHO, E. coli, Vero, yeast, NS0, Human, MDCK, Sf9 & AcNPV, Hi5 & AcNPV, plasmid, SV40LTA & EIA, etc.), and includes reagents for Proteinase K digestion, DNA binding, washing and elution, with magnetic particles for nucleic acid capture.
Technical Specifications
| Parameter | Details |
|---|---|
| Kit size / throughput | Reagents for 100 extractions |
| Sample input (typical) | 100 µL sample per extraction (procedure described: add 100 µL sample + 10 µL 5M NaCl before digestion) |
| Proteinase K digestion | Prepare Proteinase K digestion solution in Proteinase K Buffer. For samples with 0–100 mg/mL protein add 10 µL Proteinase K per 100 µL buffer; for 100–200 mg/mL protein add 20 µL Proteinase K per 100 µL buffer. Incubate digestion at 55 °C for 60 min. |
| Binding buffer (per sample) | 200 µL Binding solution + 9 µL Glycogen + 0.2 µL Yeast tRNA. Note: Do not add Yeast tRNA when extracting E. coli or yeast DNA. |
| Magnetic particles usage | Add 10 µL magnetic particles per sample. Vortex particles 5 seconds to fully resuspend before use. Store magnetic particles at 2–8 °C. |
| Alcohol addition for binding | After binding buffer, add 200 µL isopropanol per sample to facilitate binding. |
| Binding incubation | Vortex tubes vertically at medium speed for 5 minutes to bind nucleic acids to magnetic particles. |
| Wash steps | Wash once with 700 µL Wash buffer A (ethanol to be added as instructed), then wash with 700 µL Wash buffer B (70% ethanol prepared separately). Spin 10 seconds between steps and use magnetic stand to separate beads. |
| Bead drying | Air-dry magnetic pellet at room temperature for 30 seconds to 3 minutes (environment dependent) to remove residual ethanol; avoid over-drying which hinders resuspension. |
| Elution | Add 50–100 µL pre-warmed (70 °C) Elution buffer, vortex 5 seconds, incubate at 70 °C for 7 minutes with intermittent vortexing. Centrifuge 1 min, separate beads on magnetic stand and transfer eluate. |
| Typical centrifugation | Quick spins of 10 seconds are used throughout to collect liquids from caps/walls; final centrifuge after elution 1 minute. |
| Reagent storage conditions | Wash Buffer A: room temperature (add 40 mL anhydrous ethanol before use); Binding solution / Elution buffer / Dilution buffer / Proteinase K Buffer: room temperature (as labeled); Magnetic particles: 2–8 °C; Proteinase K: (no explicit temperature on label in table) - recommended follow label; Glycogen: -20 °C; Yeast tRNA: per label. |
| Shelf life | Kit components can be stored under their appropriate conditions for up to 24 months (check label expiration dates). |
| pH requirement | Adjust sample pH to neutral (pH 6.0–8.0) with 1M HCl or 1M NaOH prior to extraction if sample pH <5 or >9. |
| Controls recommended | Use Negative Control Sample (NCS) processed in parallel and Sample Extraction Recovery Control (ERC) spiked at 2–10× the amount quantified in the unspiked sample to monitor extraction recovery and accuracy. |
Features
- Magnetic-particle based extraction: Magnetic particle separation technology enables efficient capture and reproducible recovery of trace residual DNA from complex biological matrices.
- Broad host-cell compatibility: Validated for multiple host cells including CHO, E. coli, Vero, yeast, NS0, Human, MDCK, Sf9 & AcNPV, Hi5 & AcNPV, plasmid, SV40LTA & EIA.
- Manual and automated workflows: Compatible with manual preparation or automated extraction using the rHCDpurify instrument.
- Optimized for downstream quantitation: Extracted DNA is suitable for SHENTEK® host cell DNA quantitation and size analysis kits and analysis on real-time PCR systems.
- Pre-configured reagents for 100 extractions: Complete reagent set supplied for 100 DNA extractions, with specific buffers and additives (Glycogen, tRNA) for improved recovery of low-abundance DNA.
Applications
- Residual host cell DNA extraction and purification from biologics and process intermediates
- Sample preparation for host-cell DNA quantitation by qPCR
- Process monitoring for biologics and vaccine manufacturing
- Size analysis and downstream molecular assays of residual DNA
- Extraction from diverse matrices including upstream and downstream process samples
Kit Contents
Wash Buffer A (NND014) 30 mL × 1 bottle (add 40 mL anhydrous ethanol before first use)Room temperature
Binding Solution (NND016) 20 mL × 1 bottleRoom temperature
Elution Buffer (NND018) 10 mL × 1 bottleRoom temperature
Dilution Buffer (NND021) 10 mL × 1 bottleRoom temperature
Proteinase K Buffer (NND026) 10 mL × 1 bottleRoom temperature
Magnetic particles (NND031) 1 mL × 1 tube2–8 °C
Proteinase K (NND023) 500 µL × 2 tubesFollow label (store frozen or per label); prepare fresh digestion solution as instructed
Glycogen (NND035) 500 µL × 2 tubes-20 °C
Yeast tRNA (NND037) 50 µL × 1 tubeFollow label (store frozen or per label)
* Total: 100 extractions | Method: Magnetic particle separation (sample extraction)
Attention
• Read Material Safety Data Sheets (MSDS) and follow handling instructions; wear protective eyewear, clothing, and gloves.
• Add 40 mL anhydrous ethanol to Wash Buffer A before first use; prepare 70% Wash Buffer B separately and label.
• Prepare and use 100% isopropanol for binding step as instructed.
• Ensure reagents are clear before use; if cloudy, heat at 37 °C until clear and mix well.
• Adjust sample pH to neutral (pH 6.0–8.0) if outside range to avoid affecting extraction.
• Do not add Yeast tRNA when extracting E. coli or yeast DNA.
• Avoid over-drying magnetic pellet; air-dry 30 seconds to 3 minutes depending on environment.
• Perform downstream assays on the same day after nucleic acid extraction for accurate results.
• Store magnetic particles at 2–8 °C; storage below -18 °C may degrade performance.
• Use low-retention filter tips and regularly calibrate pipettes to ensure accurate spiking and aspiration.
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 sample volume is recommended per extraction?
The protocol describes using 100 µL of sample per extraction. Adjustments may be made with proportional reagent preparation; follow kit guidelines for dilution and recovery controls.
Q2: Can I use this kit for E. coli and yeast DNA extraction?
Yes. The kit supports E. coli and yeast DNA extraction; note that Yeast tRNA should not be added to the Binding buffer when extracting E. coli or yeast DNA.
Q3: What are the recommended incubation conditions for Proteinase K digestion?
Prepare Proteinase K digestion solution according to sample protein concentration and incubate at 55 °C for 60 minutes as described in the protocol.
Q4: How should I store the magnetic particles and other reagents?
Store magnetic particles at 2–8 °C. Glycogen is stored at -20 °C. Other liquid buffers are stored at room temperature as labeled. The kit components can be stored under appropriate conditions for up to 24 months; always check expiration dates on labels.
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.
