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Product Description
MycoSHENTEK® Mycobacteria DNA Extraction Kit is designed for extraction of trace Mycobacteria DNA from a variety of biologic sample types. The kit supports direct extraction from sample volumes ≤400 μL or concentration of larger volumes by centrifugation prior to extraction. It is compatible with manual magnetic-particle based workflows and automated extraction using the rHCDpurify/rDNApurify instrument. The kit workflow includes pretreatment, Proteinase K digestion, magnetic particle binding (with isopropanol), two wash steps (ethanol-based), and heat elution to produce PCR-ready DNA (minimum recommended eluate ≥40 μL). The kit is supplied for research use only and is intended for downstream molecular detection (e.g., real-time PCR) with MycoSHENTEK detection reagents or other PCR systems.
Technical Specifications
| Parameter | Details |
|---|---|
| Product number / kit size | Product No.: 1503601; reagents for 50 extractions |
| Intended use | Extraction of Mycobacteria DNA from cell-free and cell culture samples for downstream molecular detection (research use only) |
| Sample input volume | Recommended ≤400 μL per sample; if >400 μL concentrate to ≈400 μL by centrifugation |
| Proteinase K | Add 20 μL Proteinase K per sample during digestion |
| Lysis | Add 100 μL Lysis buffer; incubate at 55 °C for 60 min (digestion) |
| Binding | Add 200 μL Binding solution + 200 μL isopropanol + 50 μL Magnetic particles per sample; shake 5 min for binding |
| Washes | Wash 1: 700 μL Wash buffer A (ethanol added before use); Wash 2: 700 μL Wash buffer B (70% ethanol) |
| Elution | Add 50 μL Elution buffer; incubate at 70 °C for 7 min; recover at least 40 μL eluate |
| Automated run time | rHCDpurify / rDNApurify program: ~50 minutes (plate-based automated run) |
| Centrifugation speeds / times | Cell-free sample concentration: 16000×g for 15 min; Cell pelleting: 72×g for 5 min; Debris precipitation: 450×g for 10–15 min; brief spins (3 s) used frequently |
| Magnetic particle separation time | Separation typically 3–5 min |
| Air-dry time (post-wash) | 30 s to 3 min depending on environment; avoid over-drying |
| Storage stability | Kit components storable under indicated conditions for up to 24 months |
Features
- Trace DNA extraction: Optimized to extract low-abundance (trace) Mycobacteria DNA from various biologic matrices.
- Magnetic bead-based purification: Magnetic particle chemistry provides efficient capture and wash-based purification compatible with manual and automated workflows.
- Manual and automated workflows: Validated for manual tube-based extraction and automated plate-based extraction using rHCDpurify / rDNApurify instruments.
- PCR-ready eluate: Produces a concentrated eluate (recommended ≥40 μL) suitable for downstream real-time PCR detection.
- Room-stable major reagents: Most reagents are stored at room temperature; selected reagents (e.g., cell lysis/pretreatment/magnetic particles) refrigerated as indicated.
- Complete workflow reagents included: Includes lysis, binding, wash, elution buffers, proteinase K, magnetic particles and pretreatment reagents for sample types.
Applications
- Extraction of Mycobacteria DNA from cell-free biologics (e.g., serum, plasma, cell-free supernatants)
- Extraction of Mycobacteria DNA from cell culture samples and cell pellets
- Sample preparation for downstream molecular assays including real-time PCR/qPCR
- Research applications and in-process quality control testing of biologics (Research Use Only)
Kit Contents
Lysis buffer (NND028)Room temperature; 5 mL × 1 bottle
Binding solution (NND017)Room temperature; 10 mL × 1 bottle
Wash buffer A (NND015) — add 20 mL anhydrous ethanol before useRoom temperature; 15 mL × 1 bottle
Elution buffer (NND019)Room temperature; 5 mL × 1 bottle
Dilution buffer (NND022)Room temperature; 5 mL × 1 bottle
MB Cell lysis buffer (NND039)2–8 °C; 1.25 mL × 2 tubes
5M NaCl (NND040)Follow label; 500 μL × 1 tube
Pretreatment buffer (NND041)2–8 °C; 1.25 mL × 2 tubes
Magnetic particles (NND034)Follow label (equilibrate to room temp before use); 1.25 mL × 2 tubes
Proteinase K (NND024)Follow label; 1 mL × 1 tube
* Total: 50 extractions | Method: Magnetic particle-based DNA extraction (manual or automated)
Attention
• For research use only. Read Material Safety Data Sheets (MSDS) and use appropriate PPE (gloves, lab coat, eye protection, mask).
• Use separate, dedicated laboratory areas and equipment for reagent preparation, sample preparation, amplification, and post-amplification to minimize contamination.
• Ensure ambient temperature ≥22 °C before starting experiments when recommended.
• Balance and vortex magnetic particles before use; centrifuge tubes briefly (3 s) when instructed to collect liquid from caps/walls.
• Prepare Wash buffer A by adding 20 mL anhydrous ethanol before first use; prepare Wash buffer B as 70% ethanol separately.
• Do not over-dry the magnetic particle pellet after ethanol wash; over-drying may make resuspension difficult.
• Perform mycobacterial DNA detection assays the same day as extraction to ensure accuracy.
• For automated runs: perform UV sterilization of the instrument for at least 15 min and clean internal walls with 75% alcohol wipes before and after runs; minimum interval between extractions is 30 min.
• After automated extraction, immediately transfer eluate to a new tube and ensure ≥40 μL is recovered for downstream assays.
• Dispose of used tips and liquid waste according to laboratory biohazard and local regulations.
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 types are compatible with this kit?
The kit is validated for cell-free samples (e.g., serum, plasma, supernatants) and cell culture samples (cell pellets/resuspensions). Samples ≤400 μL can be processed directly; larger volumes should be concentrated to ≈400 μL before extraction.
Q2: What is the recommended elution volume and expected yield for downstream PCR?
Standard elution uses 50 μL Elution buffer with incubation at 70 °C for 7 minutes. Recover at least 40 μL eluate for downstream PCR/qPCR applications.
Q3: Can I run the kit on an automated instrument?
Yes. The kit is compatible with rHCDpurify / rDNApurify automated plate-based instruments. The automated program run time is approximately 50 minutes for a typical plate protocol.
Q4: How should magnetic particles be handled before use?
Equilibrate magnetic particles to room temperature for 10 minutes and vortex thoroughly to fully resuspend particles before aliquoting to samples to ensure consistent binding performance.
Research Use Only
✔
Research Use Only (RUO) — For research purposes; not for diagnostic use – 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.
