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| Product Name | Magnetic Bead-Based Pathogen DNA/RNA Extraction Kit |
| Catalog No. | DREK-0009 |
| Description | A magnetic bead-based total nucleic acid extraction kit designed for the simultaneous extraction of pathogen DNA and RNA from a wide variety of clinical and environmental sample matrices. The kit employs a universal lysis buffer with mechanical disruption capability for difficult-to-lyse organisms including Gram-positive bacteria, mycobacteria, fungi, and protozoan parasites. The optimized bead chemistry, combined with carrier RNA supplementation, ensures high-efficiency recovery of low-abundance pathogen targets in the presence of high host background nucleic acids. |
| Intended Use | Simultaneous extraction of microbial and viral DNA and RNA from diverse clinical specimens, environmental samples, food matrices, and veterinary specimens for comprehensive molecular pathogen detection, microbiome analysis, and antimicrobial resistance gene profiling. |
| Principle / Technology | Samples are subjected to a combination of chemical lysis (guanidine isothiocyanate, SDS, and lysozyme) and optional mechanical disruption (bead beating with ceramic or glass beads). Released nucleic acids are captured by silica-coated magnetic beads under optimized binding conditions that support both DNA and RNA binding. Carrier RNA is included to improve recovery of low-titer targets. After stringent washing, purified total nucleic acids are eluted and ready for downstream detection. |
| Detection Method | qPCR and RT-qPCR with pathogen-specific and broad-range primer sets (16S rRNA, ITS, 18S rRNA); fluorometric quantitation (Qubit for DNA, Qubit RNA for RNA); Bioanalyzer for fragment analysis; metagenomic NGS for comprehensive pathogen profiling and antimicrobial resistance gene detection |
| Sample Type | Clinical specimens: whole blood, plasma, serum, sputum, bronchoalveolar lavage, cerebrospinal fluid, urine, stool, pus, wound swabs, tissue biopsies; environmental: soil, water, air filters, surface swabs; food: homogenates, rinsates, enrichment broths; veterinary: whole blood, tissue, fecal samples from livestock, companion animals, and wildlife |
| Performance Range / Specifications | Input volume: 100-500 µL liquid sample or 25-100 mg solid sample; DNA/RNA recovery: ≥70% recovery of bacterial and viral reference standards at 10³ CFU or copies/mL; A260/A280: 1.7-2.0; total nucleic acid yield: 100 ng-20 µg (sample-dependent); simultaneous recovery confirmed by separate DNA and RNA detection assays |
| Sensitivity / LOD | Detection of 10-50 CFU bacteria per mL and 50-200 viral copies per mL from liquid samples; effective lysis of Gram-positive, Gram-negative, acid-fast, and fungal organisms confirmed by broad-range PCR; carrier RNA enables detection of <10 copies of RNA virus per reaction |
| Specificity | Broad-spectrum pathogen nucleic acid recovery verified across bacterial, viral, fungal, and parasitic species panels; minimal bias toward Gram-negative organisms; unbiased co-recovery of DNA and RNA verified by spike-in experiments; no PCR inhibition in 1:5 diluted extracts from complex matrices including stool and soil |
| Reaction Conditions / Protocol | Transfer sample to lysis tube containing lysis buffer and optional ceramic beads; vortex or bead-beat for 5-15 minutes; add proteinase K; incubate at 56°C for 15 minutes; add binding buffer, carrier RNA, and isopropanol; add magnetic bead suspension; incubate 10 minutes; separate beads; wash once with wash buffer 1; wash twice with wash buffer 2 (ethanol-based); dry beads; elute in 50-100 µL nuclease-free water at 56°C for 5 minutes; total time: 60-90 minutes |
| Components / Formulation | Lysis buffer (guanidine isothiocyanate, Tris-HCl, EDTA, SDS, Triton X-100), lysozyme (lyophilized), carrier RNA (poly-A, lyophilized), proteinase K solution (20 mg/mL), binding buffer (guanidine hydrochloride, Tris-HCl, sodium acetate, isopropanol), magnetic silica bead suspension (50 mg/mL), wash buffer 1 concentrate, wash buffer 2 concentrate, nuclease-free water, ceramic disruption beads (0.1 mm and 0.5 mm mix), magnetic separation rack |
| Storage Conditions | Lysis buffer and wash buffers at room temperature; lysozyme at -20°C lyophilized, 2-8°C after reconstitution; carrier RNA at -20°C; proteinase K at 2-8°C; magnetic bead suspension at 2-8°C; ceramic beads at room temperature |
| Shelf Life | 18 months from date of manufacture for buffers and beads; lysozyme 24 months lyophilized at -20°C, 4 weeks at 2-8°C after reconstitution; carrier RNA 12 months at -20°C; proteinase K 12 months at 2-8°C after opening |
| Package Specifications | 50 preparations and 200 preparations; includes ceramic disruption beads, lysozyme, carrier RNA, and all required reagents; magnetic rack sold separately |
| Product Form | Liquid buffers; silica-coated magnetic bead suspension; lyophilized lysozyme and carrier RNA; ceramic disruption beads; proteinase K solution |
| Quality Control | Each lot validated with: E. coli (Gram-negative), S. aureus (Gram-positive), C. albicans (fungal), and MS2 bacteriophage (RNA virus) spike-in panel: ≥70% recovery for all targets by qPCR/RT-qPCR; gDNA-free confirmed for RNA targets; PCR inhibitor test negative for 7 common inhibitors; RNase/DNase-free certification |
| Key Features | Universal lysis system effective against all pathogen classes; simultaneous DNA and RNA recovery; ceramic bead beating option for refractory organisms; carrier RNA for low-titer target enhancement; magnetic bead technology compatible with automation; no phenol-chloroform extractions |
| Purity | A260/A280: 1.7-2.0 for mixed nucleic acid extracts; no detectable protein contamination by Bradford assay |
| Concentration | Elution volume 50-100 µL; total nucleic acid concentration variable by sample type; typical concentration 2-50 ng/µL |
| Activity / Unit Definition | Not applicable for nucleic acid extraction kits |
| Molecular Weight | Not applicable for heterogeneous nucleic acid extracts containing both DNA and RNA of diverse sizes and origins |
| Source / Origin | Silica-coated magnetic beads from synthetic process; lysozyme from hen egg white (chromatographically purified); carrier RNA is synthetic poly-A; proteinase K expressed recombinantly; ceramic beads are yttria-stabilized zirconia |
| pH Range / Optimal pH | Lysis buffer pH 6.0-6.5; binding buffer pH 5.0-6.0; wash buffers pH 6.5-7.5; elution water pH 6.5-7.5 |
| Shipping Conditions | Ambient temperature for buffers and ceramic beads; lysozyme and carrier RNA shipped on dry ice; proteinase K shipped with cold packs |
| Expiration Date / Stability | 18 months shelf life; real-time stability confirmed at 18 months with pathogen panel testing; lysozyme activity maintained >80% at recommended storage; carrier RNA integrity verified per lot by gel electrophoresis |
| Regulatory / Compliance | For research use only; suitable for pathogen surveillance, environmental monitoring, and food safety testing research; not for clinical diagnostic use; appropriate biosafety level containment required for pathogenic organisms |
| Compatibility | Extracted nucleic acids suitable for qPCR, RT-qPCR, digital PCR, isothermal amplification (LAMP, RPA), microarray, and metagenomic NGS; compatible with open-platform magnetic bead handlers including KingFisher systems; validated with major qPCR master mixes |
| Recommended Buffer System | Guanidine isothiocyanate-SDS-Triton lysis buffer; lysozyme supplementation for peptidoglycan digestion; guanidine hydrochloride binding buffer; nuclease-free water elution |
| Application Notes / Precautions | For samples with high PCR inhibitor content (stool, soil, plant material), include an additional wash step; for mycobacteria and fungal elements, include bead beating at maximum speed; for precious low-volume samples, reduce elution volume to 20 µL; use nuclease-free consumables throughout; handle carrier RNA with care to avoid RNase contamination; pre-aliquot carrier RNA to minimize freeze-thaw cycles; run extraction-negative controls with each batch |
| Batch-to-Batch Consistency | Between-lot recovery CV <18% for bacterial and viral spike-in targets; binding capacity ≥20 µg total nucleic acid per preparation; lysozyme specific activity maintained within ±15% between batches; carrier RNA size distribution verified per lot; magnetic bead lot traceability maintained |
For research use only, not for clinical use.
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