0 item(s)
View cart
You have no items in your shopping cart.
For Research Use Only (RUO)
$3,708.00
Sku: SK030307S-P
Product Description
SHENTEK® Residual HPV18 E6/E7 DNA Size Analysis Kit is a real-time PCR (qPCR) kit designed for rapid and specific quantitation of residual HPV18 E6 and E7 DNA fragments of different sizes (from HeLa cell origin) across biopharmaceutical sample types (in-process to final products). The kit amplifies four fragment lengths (E6: 100 bp and 288 bp; E7: 110 bp and 240 bp) using FAM-labeled target probes and includes an internal positive control (IPC, VIC) for inhibition monitoring. The kit includes HPV E6/E7 DNA Control for standard curve generation and is intended for research use only. For sample extraction instructions refer to the SHENTEK® Residual Host Cell DNA Sample Preparation Kit User Guide (Product No. 1104191).
Technical Specifications
| Parameter | Details |
|---|---|
| Target genes / fragments | HPV18 E6 (100 bp, 288 bp) and E7 (110 bp, 240 bp) |
| Detection chemistry / channels | FAM-labeled probes for target fragments; VIC-labeled probe for IPC; ROX as passive reference (recommended) |
| Reaction volume | 30 μL total reaction volume (20 μL qPCR mix + 10 μL sample or standard) |
| qPCR mix composition (per reaction) | qPCR Reaction Buffer 17 μL + primer&probe mix 3 μL (per target assay); IPC qPCR mix: qPCR Reaction Buffer 15.9 μL, IPC MIX 1.3 μL, ddH2O 2.8 μL (per 20 μL qPCR MIX) |
| Cycling conditions | Activation 95 °C 10:00 (1 cycle); 40 cycles of 95 °C 00:15 and 60 °C 01:00 (fluorescence read during annealing/extension) |
| Standard curve / dynamic range (provided example) | Standard curve points shown: 1×10^8 to 1×10^2 copies/μL (ST0 = 1×10^8, ST1 = 1×10^7 ... ST6 = 1×10^2); at least five concentrations recommended |
| Replicates | Triplicates recommended for standards, controls and test samples |
| Controls included / required | HPV E6/E7 DNA Control (quantified) for standard curve; NTC (no template control); NCS (negative control sample); IPC for inhibition monitoring |
| Input/sample volume | 10 μL purified sample or 10 μL standard per reaction |
| Kit throughput | Reagents for 4 × 100 reactions (as labeled on product) |
| Shelf life | Up to 24 months when stored at –20 °C (check label expiration date) |
| LOD / LOQ | Not specified in the user guide; recommended to determine during in-house validation |
| Inhibition acceptance criterion | Mean Ct of IPC for sample should be within ±1.0 of the NCS Ct-IPC; significant increase suggests inhibition and sample recovery testing is recommended |
| Compatible instruments | SHENTEK-96S, ABI 7500, Bio-Rad CFX96, LineGene 9600plus and other real-time PCR systems (user must configure detectors/filters accordingly) |
Features
- Fragment-size specific quantitation: Designed to amplify and quantify four distinct HPV18 E6/E7 fragment sizes (E6:100 bp & 288 bp; E7:110 bp & 240 bp) to determine size distribution of residual DNA.
- Real-time qPCR with internal control: Uses FAM-labeled probes for targets and VIC-labeled IPC to monitor inhibition and assay performance.
- Pre-formulated primer & probe mixes: Supplied primer & probe mixes for each fragment simplify setup and reduce pipetting errors; probes are light-sensitive (protect from light).
- Ready-to-use standards: Includes quantified HPV E6/E7 DNA Control and detailed serial dilution protocol for building standard curves and absolute quantitation.
- Validated workflow guidance: Provides recommended plate layouts, qPCR conditions, and data analysis settings for ABI7500 (can be adapted to other instruments).
Applications
- Quantitation of residual HPV18 E6/E7 DNA (HeLa origin) in biopharmaceutical production
- In-process monitoring and final product quality control for residual host cell DNA (HPV18-specific)
- Method validation and assay development for residual DNA size-distribution analysis
- Sample inhibition assessment and recovery evaluation using IPC
Kit Contents
IPC MIX150 μL ×1 tube; –20 °C, protect from light
E6-100 primer & probe MIX300 μL ×1 tube; –20 °C, protect from light
E6-288 primer & probe MIX300 μL ×1 tube; –20 °C, protect from light
E7-110 primer & probe MIX300 μL ×1 tube; –20 °C, protect from light
E7-240 primer & probe MIX300 μL ×1 tube; –20 °C, protect from light
qPCR Reaction Buffer (NNB001)850 μL × multiple tubes (as supplied); –20 °C
DNA Dilution Buffer (DDB, NND001)1.5 mL ×2 tubes; store at 2–8 °C (if cloudy, heat to 37 °C to clarify)
HPV E6/E7 DNA Control (NNA030)50 μL ×1 tube; –20 °C (check label for concentration before dilution)
* Total: 4 × 100 reactions | Method: qPCR (real-time PCR, absolute quantitation using standard curve)
Attention
• For Research Use Only (not for diagnostic or clinical use).
• Read Material Safety Data Sheets (MSDS) and follow handling instructions; wear protective eyewear, mask, lab coat and gloves.
• Protect primer & probe mixes and IPC MIX from light; store kit components at –20 °C and check expiration date.
• Decontaminate work surfaces, pipettes and tubes (UV 30 minutes recommended) and use dedicated pre- and post-PCR areas to minimize contamination.
• Prepare and run NTC, NCS and IPC controls as instructed. NCS must be processed the same as test samples.
• If IPC Ct for a sample is > ±1.0 compared to NCS IPC Ct, the sample may contain inhibitors; perform recovery testing or further sample cleanup.
• Unused DNA Dilution Buffer (DDB) should be stored at 2–8 °C; if cloudy, warm to 37 °C to clarify.
• Follow instrument-specific software settings and thresholds; example settings provided for ABI 7500 (Manual Ct, threshold 0.02, automatic baseline).
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 expected assay runtime?
The qPCR program provided runs activation 95 °C for 10 min plus 40 cycles of 95 °C for 15 s and 60 °C for 60 s, resulting in an approximate total runtime of ~1 hour per run.
Q2: How should I prepare the standard curve?
Thaw the supplied HPV E6/E7 DNA Control, dilute to prepare ST0 at 1×10^8 copies/μL, then perform serial 1:10 dilutions to generate ST1–ST6 (1×10^7 to 1×10^2 copies/μL). Include at least five concentrations across the dynamic range and run in triplicate.
Q3: What should I do if the IPC indicates inhibition?
If sample mean Ct-IPC is significantly higher than NCS Ct-IPC (>±1.0), the sample may be inhibitory. Perform recovery testing (ERC) in the same assay, consider additional sample cleanup or dilution, and re-test.
Q4: What is the assay sensitivity (LOD/LOQ)?
LOD/LOQ are not specified in the user guide. Determine LOD/LOQ, precision and accuracy during in-house validation using the provided standard curve and appropriate validation procedures.
Research Use Only
✔
Research Use Only (RUO). Products are intended for research use only. – 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.
