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For Research Use Only (RUO)
$1,751.00
Sku: 1801940
Product Description
The SHENTEK 10‑Species Cell Line Authentication Kit is a multiplex PCR‑based assay for rapid species verification and cross‑contamination detection of commonly used cell lines in biologics manufacturing. The kit targets mitochondrial genes (Cytochrome Oxidase I (COI), Cytochrome B (CytB), and Cytochrome Oxidase II (COII)) using species‑specific primers that generate distinct amplicon sizes for ten species in a single reaction. Results are analyzed by horizontal agarose gel electrophoresis; species identification and mixed‑species contamination are inferred from the number and sizes of amplicons. The assay has been validated to detect contamination down to ≤1% and includes reagents sufficient for 50 reactions.
Technical Specifications
| Parameter | Details |
|---|---|
| Intended use / Compliance | Research Use Only (RUO); for cell line species identification and contamination detection |
| Detection limit | Validated to detect cross‑contamination at ≤1% (by proportion of species in a mixed sample) |
| Target genes | Mitochondrial genes: Cytochrome Oxidase I (COI), Cytochrome B (CytB), Cytochrome Oxidase II (COII) |
| Species detected (amplicon sizes) | Bos taurus (Cow) 93 bp; Mus musculus (Mouse) 147 bp; Canis familiaris (Dog) 172 bp; Rattus norvegicus (Rat) 200 bp; Cercopithecus aethiops (African green monkey) 256 bp; Macaca mulatta (Rhesus monkey) 287 bp; Cricetulus griseus (Chinese hamster) 315 bp; Felis catus (Cat) 355 bp; Homo sapiens (Human) 394 bp; Sus scrofa (Pig) 464 bp |
| Sample input | Recommended starting material: 5×10^5 freshly cultured cells per extraction |
| Sample extraction conditions | Add 100 µL Cell Extraction Buffer; incubate 60 °C for 1 hour, then 95 °C for 10 minutes; extracts may be stored at -20 °C for up to 1 month |
| Reaction format and volumes | 25 µL total reaction: 17 µL PCR MIX (12.5 µL 2× Multiplex PCR Buffer + 4.5 µL DNA Sizing Primers MIX) + 8 µL template (extract/controls) |
| Kit capacity | Reagents for 50 reactions (as supplied) |
| Thermal cycling program | 1) 95 °C 5 min (1 cycle); 2) [95 °C 30 s; 62 °C 3 min; 68 °C 30 s] for 25–30 cycles (28 cycles recommended for pilot study); 3) 68 °C 30 min; 4) 4 °C hold |
| Agarose electrophoresis conditions | Use 2% agarose gel in 1× TBE; run at constant 80–110 V until dye front ~60 mm; recommended loading 6–10 µL per well; include DL500 DNA marker |
| DNA Sizing Ladder | Lane 11 (DNA sizing ladder) contains 10 distinguishable bands corresponding to the 10 species; ensure clear visualization for reliable species calling |
| Storage of kit components | Kit components can be stored under appropriate conditions for up to 24 months (individual reagent storage listed in kit contents) |
| Recommended replicates | Analyze each test sample in two independent replicates to ensure reliability |
Features
- Multiplex PCR for simultaneous detection: Detects and differentiates 10 species in a single PCR reaction using species‑specific primers that produce distinct amplicon sizes.
- Mitochondrial gene targets: Targets mitochondrial genes (COI, CytB, COII) for robust species discrimination across common cell lines.
- High sensitivity for contamination detection: Validated to detect contaminating species present at or below 1% of the sample.
- Gel‑based readout: Simple analysis by horizontal agarose electrophoresis — no sequencing required; identification based on band size and pattern.
- Optimized workflow: Includes cell extraction buffer and pre‑mixed primer and template reagents; protocol provides recommended PCR cycling and gel conditions for reproducible results.
Applications
- Cell line species verification
- Cross‑contamination detection in cell cultures
- Quality control for biologics manufacturing
- Research use for cell authentication in academic and industrial labs
Kit Contents
DNA Sizing Templates MIX (NNA048) 25 µL × 2 tubes-20 °C
Cell Extraction Buffer (NNB015) 1.25 mL × 4 tubesStore as labeled; kit components stable up to 24 months
2× Multiplex PCR Buffer (NNB016) 320 µL × 2 tubesStore as labeled; kit components stable up to 24 months
DNA Sizing Primers MIX (NNC082) 120 µL × 2 tubesStore as labeled; kit components stable up to 24 months
DEPC‑treated H2O (NND052) 1 mL × 2 tubesStore as labeled; kit components stable up to 24 months
DL500 DNA Marker, 1 tubeStore at 2–8 °C after thawing; aliquot to avoid repeated freeze–thaw
Complementary reagents (sold separately): 6× Loading Buffer (Product No. 1801942) and 10× TBE Buffer (Product No. 1801943)Refer to individual product labels
* Total: 50 reactions | Method: Multiplex PCR
Attention
• For Research Use Only (RUO). Not for diagnostic use.
• Read Material Safety Data Sheets (MSDS) and follow handling instructions; use appropriate PPE (gloves, mask, eyewear, lab coat).
• Avoid cross‑contamination: use physically separated areas for sample preparation, reagent setup (negative area), positive control handling, and amplification; if separate rooms are not available, clearly demarcate areas.
• Perform a pilot study to verify PCR program settings and electrophoresis conditions before routine use.
• Open PCR tubes/caps only in the electrophoresis area; reseal opened amplification products if not immediately loaded to avoid contamination.
• Avoid repeated freeze–thaw cycles of reagents and extracts; DNA extracts may be stored at -20 °C for up to 1 month.
• Include appropriate controls: No Template Control (NTC), Negative Control Sample (NCS), and DNA sizing ladder; NTC and NCS should show no amplification.
• Analyze each test sample in two independent replicates to ensure reliability.
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 minimum proportion of contaminating species that can be detected?
The assay has been validated to detect contaminating species at or below 1% of the total sample.
Q2: How much sample is required for extraction?
The recommended starting material is 5×10^5 freshly cultured cells per extraction; follow the extraction protocol (100 µL Cell Extraction Buffer, 60 °C for 1 hour, 95 °C for 10 minutes).
Q3: How are species identified from the gel?
Species are identified by comparing amplicon band sizes to the DNA sizing ladder. Distinct band sizes (provided in the kit documentation) correspond to specific species; multiple bands indicate mixed‑species contamination.
Q4: How many reactions are included in the kit and what is the reaction format?
The kit provides reagents for 50 reactions. Standard reaction volume is 25 µL (17 µL PCR MIX + 8 µL template).
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.
