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Product Description
MycoSHENTEK® Mycobacteria DNA Detection Kit is used, together with MycoSHENTEK® Mycobacteria DNA Extraction Kit, to qualitatively determine whether there is Mycobacteria contamination in cells, cell culture-derived biologicals, and similar materials using real-time qPCR technology. The integrated system has been validated with a limit of detection of 10–100 CFU/mL and can detect more than 100 different Mycobacteria species. The assay chemistry includes dUTP and is compatible with UNG (uracil-N-glycosylase) treatment to prevent carry-over contamination. An Internal Control (IC) is provided to monitor PCR inhibition and/or extraction efficiency. The kit is supplied with positive and negative controls and is intended for research use only.
Technical Specifications
| Parameter | Details |
|---|---|
| Product Number | 1503602 |
| Kit Format / Reactions | Reagents for 50 Reactions |
| Intended use | Qualitative detection of Mycobacteria contamination in cells and cell culture-derived biologicals |
| Limit of Detection (LOD) | Validated at 10–100 CFU/mL |
| Species coverage | Validated to detect >100 Mycobacteria species |
| Assay type | Real-time qPCR (probe-based) |
| Total reaction volume | 30 µL (10 µL qPCR MIX + 20 µL sample/control) |
| qPCR MIX composition (per reaction) | MB qPCR Reaction Buffer 8 µL; MB Primer & Probe MIX 1.5 µL; Internal Control (IC) 0.5 µL; total 10 µL. UNG optional (0.1 U per reaction). |
| Cycling conditions | UNG 25 °C 10:00 (1 cycle); Activation 95 °C 10:00 (1 cycle); 45 cycles of Denaturation 95 °C 00:15, Annealing 60 °C 00:30, Extension 72 °C 01:30 (read fluorescence during extension). |
| Controls | Positive Control (PC), No Template Control (NTC), Negative Control Sample (NCS), Internal Control (IC) |
| Acceptance criteria (Ct thresholds) | Typical interpretation: FAM positive cutoff Ct <40.00 (positive if at least one well Ct<40 and normal amplification); VIC (IC) expected Ct <35.00 for normal amplification; PC Ct <35.00. NTC FAM Ct ≥40.00 or no amplification. |
| Compatible instruments | SHENTEK-96S, Bio-Rad CFX96, Applied Biosystems 7500, and other real-time PCR systems supporting probe detection |
| Storage stability | Kit components can be stored under appropriate conditions for up to 24 months (see individual component storage) |
Features
- High sensitivity: Validated limit of detection down to 10–100 CFU/mL for broad Mycobacteria detection.
- Broad species coverage: Designed and validated to detect over 100 different Mycobacteria species.
- Carry-over contamination control: Chemistry includes dUTP and is compatible with UNG treatment to prevent and eliminate carry-over contamination.
- Internal control (IC): IC monitors PCR inhibition and/or extraction efficiency; can be added pre-extraction or to the PCR mix.
- Pre-validated workflow: Includes positive and negative controls and recommended plate layouts and acceptance criteria for straightforward result interpretation.
- Compatibility: Works with common real-time PCR instruments (e.g., SHENTEK-96S, CFX96, 7500).
Applications
- Qualitative detection of Mycobacteria contamination in cell lines
- Screening of cell culture-derived biologicals for Mycobacteria contamination
- Routine mycoplasma/contaminant monitoring workflows in research laboratories (Mycobacteria-specific)
- Research studies requiring sensitive detection of Mycobacteria DNA
Kit Contents
MB qPCR Reaction Buffer (NNB011)-20 °C, protect from light; 400 µL × 1
MB Primer & Probe MIX (NNC035)2–8 °C until use or -20 °C; 75 µL × 1
MB Internal Control (IC) (NNA037)2–8 °C or -20 °C; 600 µL × 1
MB Positive Control (PC) (NNA041)-20 °C; 500 µL × 1
DNA Dilution Buffer (DDB) (NND001)2–8 °C or room temperature per label; 1.5 mL × 1
* Total: 50 reactions | Method: qPCR
Attention
• For Research Use Only (RUO) — not for diagnostic use.
• Read Material Safety Data Sheets (MSDS) and follow handling instructions; wear protective eyewear, mask, clothing and gloves.
• Prevent contamination: use UV decontamination (30 minutes) and 75% ethanol to clean surfaces and equipment before setup.
• Thaw reagents at 2–8 °C or on ice, vortex and briefly centrifuge before use.
• Use UNG enzyme optionally to reduce carry-over contamination (check effective amount before use).
• If VIC (IC) signal is inhibited, repeat sample preparation or remove inhibitors and repeat the assay.
• Follow plate layout and include PC, NTC and NCS as described; use duplicates for samples and controls.
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 limit of detection (LOD) for this kit?
The integrated system has been validated with a limit of detection of approximately 10–100 CFU/mL.
Q2: Can the kit detect multiple Mycobacteria species?
Yes. The assay has been validated to detect more than 100 different Mycobacteria species.
Q3: Is an Internal Control provided and how should it be used?
An Internal Control (IC) is provided. It can be added during sample extraction to monitor extraction efficiency or added to the PCR mix to monitor PCR inhibition. If IC was added during extraction, replace IC volume with DNA dilution buffer when preparing the qPCR mix.
Q4: What are the recommended instruments and reaction setup?
The kit is compatible with common real-time PCR instruments (e.g., SHENTEK-96S, CFX96, 7500). Prepare 30 µL reaction volumes (10 µL qPCR MIX + 20 µL sample/control) and run the recommended cycling program with UNG treatment if used.
Research Use Only
✔
For 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.
