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Product Description
SHENTEK® Residual PG13 DNA Size Analysis Kit is designed to quantify residual PG13 host-cell DNA across different fragment sizes during biopharmaceutical production (from in-process samples to final products). The kit uses a fluorescent quantitative PCR (FAM) assay to rapidly and specifically measure PG13 DNA fragments. The assay amplifies three discrete target fragment sizes (118 bp, 212 bp and 502 bp) to determine size distribution. The kit includes an internal positive control (IPC) and a PG13 DNA control for generation of absolute-quantitation standard curves. Limit of detection (LOD) reaches the femtogram (fg) level. For DNA extraction recommendations, refer to the companion SHENTEK® Residual Host Cell DNA Sample Preparation Kit (Product No. 1104191).
Technical Specifications
| Parameter | Details |
|---|---|
| Target fragment sizes | 118 bp, 212 bp, 502 bp |
| Limit of detection (LOD) | LOD reaches femtogram (fg) level |
| Standard curve concentrations (example) | ST1 = 300 pg/µL; ST2 = 30 pg/µL; ST3 = 3 pg/µL; ST4 = 0.3 pg/µL; ST5 = 0.03 pg/µL |
| Reaction volume | Total reaction volume 30 µL (20 µL qPCR MIX + 10 µL sample) |
| qPCR MIX composition (per 1 reaction) | qPCR Reaction Buffer 15.9 µL; PG13 Primer&Probe MIX (per target) 2.8 µL; IPC MIX 1.3 µL; total 20 µL |
| 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 (read fluorescence) — 40 cycles |
| Instrument/readout channels | FAM for PG13 targets; VIC for IPC; ROX used as passive reference (ABI7500 example) |
| IPC acceptance criteria | Mean Ct(IPC) for sample should be within ±1.0 of NCS Ct(IPC); a significantly higher Ct indicates possible inhibition |
| Recovery rate (ERC) | Expected ERC recovery between 50% and 150% |
| NTC and NCS criteria | NTC Ct should be ≥35 cycles or undetermined; NCS Ct should be larger than mean Ct of lowest standard and show VIC amplification |
| Kit shelf life | Components stored at –20 °C; stable for up to 24 months (check label expiration) |
| Provided reagent format | Reagents provided for 3 × 100 reactions (stated on product cover) |
Features
- Size-resolved quantitation: Three separate primer/probe assays (118 bp, 212 bp, 502 bp) enable determination of residual DNA size distribution, not just total DNA.
- High sensitivity: Assay LOD reaches femtogram (fg) levels for detection of trace residual DNA.
- Absolute quantitation: Includes PG13 DNA control and recommended serial dilution scheme to build standard curves for absolute quantitation.
- Internal control for inhibition monitoring: IPC (VIC) included to monitor PCR inhibition and assay performance on a per-sample basis.
- Instrument compatibility: Compatible with common real-time PCR platforms (e.g., SHENTEK-96S, ABI 7500, LightCycler 480).
- Ready-to-use qPCR mixes: Pre-formulated qPCR Reaction Buffer and Primer&Probe mixes reduce setup time and variability.
Applications
- Quantitation of residual PG13 host-cell DNA in biopharmaceutical process samples
- In-process monitoring of DNA fragmentation during downstream processing
- Final product quality control and release testing for residual host-cell DNA
- Method validation and process development for gene therapy/viral vector production using PG13-derived material
Kit Contents
PG13 DNA Control (NNA049) — 50 µL × 1 tube–20 °C
qPCR Reaction Buffer (NNB001) — 850 µL × 6 tubes (protect from light)–20 °C
PG13 Primer&Probe MIX-118 (NNC083) — 300 µL × 1 tube–20 °C
PG13 Primer&Probe MIX-212 (NNC084) — 300 µL × 1 tube–20 °C
PG13 Primer&Probe MIX-502 (NNC085) — 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 3 × 100 reactions (300 reactions) | Method: qPCR (fluorescent quantitative PCR, FAM)
Attention
• For research use only (RUO). Not for diagnostic or therapeutic use.
• Read Material Safety Data Sheets (MSDS) and follow handling instructions; wear appropriate PPE (gloves, lab coat, eye protection, mask).
• Work in a clean, DNase-free environment; decontaminate surfaces and equipment (UV 30 minutes and 75% ethanol recommended).
• Thaw reagents at 2–8 °C or on ice; vortex and brief-spin before use.
• Store components at –20 °C; check expiration dates on labels.
• If DNA Dilution Buffer (DDB) is cloudy or contains precipitate, heat at 37 °C until clear before use.
• Include appropriate controls: at least 5 standard points, NTC, and NCS; recommended technical triplicates for samples.
• IPC performance: sample mean Ct(IPC) should be within ±1.0 of NCS Ct(IPC); significant deviation may indicate inhibition.
• ERC (extraction recovery control) recovery should be 50%–150%; investigate out-of-range recoveries.
• NTC Ct should be ≥35 cycles or undetermined; NCS should be greater than lowest standard Ct and show VIC amplification.
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 instruments are compatible with this kit?
The kit is compatible with common real-time PCR platforms, including SHENTEK-96S, ABI 7500 series, and LightCycler 480. If using a different instrument, follow the instrument/software documentation for detector and plate setup.
Q2: How should I prepare the standard curve?
Thaw PG13 DNA Control and DDB at 2–8 °C. Dilute the DNA Control to 3000 pg/µL (ST0), then perform serial 1:10 dilutions to generate ST1–ST5 (300, 30, 3, 0.3, 0.03 pg/µL). Include at least five standards in the standard curve and run in triplicate.
Q3: What are acceptable IPC and recovery criteria?
Mean Ct(IPC) for a sample should be within ±1.0 of the NCS Ct(IPC). ERC recovery rates should fall between 50% and 150%; values outside this range warrant investigation for extraction efficiency or inhibition.
Q4: What should I do if my DNA Dilution Buffer is cloudy?
If DDB is cloudy or contains precipitate, heat the buffer at 37 °C until it becomes clear, then cool and use. Store DDB at 2–8 °C after opening or per kit instructions.
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.
