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| Product Name | Column-Based Viral DNA/RNA Extraction Kit |
| Catalog No. | DREK-0010 |
| Description | A silica membrane spin column kit for the simultaneous extraction of viral DNA and RNA from cell-free clinical specimens. The system uses chaotropic salt-driven nucleic acid binding to silica membranes in a convenient spin-column format. Carrier RNA is included in the lysis buffer formulation to enhance binding and recovery of low-concentration viral nucleic acid targets, ensuring sensitive detection in downstream amplification workflows. |
| Intended Use | Extraction of viral DNA and RNA from plasma, serum, cerebrospinal fluid, urine, nasopharyngeal swab transport medium, and cell culture supernatant for molecular diagnosis of viral infections in research settings. |
| Principle / Technology | Viral particles are lysed in a guanidine isothiocyanate-based buffer containing carrier RNA. After addition of ethanol, the lysate is passed through a silica membrane spin column where nucleic acids bind to the silica matrix. Two sequential ethanol-based wash steps remove proteins, salts, and other contaminants. Purified viral nucleic acids are eluted in a low-salt buffer. |
| Detection Method | UV spectrophotometry; qPCR and RT-qPCR with virus-specific primer/probe sets; digital PCR for absolute quantification; control template spike-in recovery monitoring |
| Sample Type | Human plasma (EDTA or citrate anticoagulated), serum, cerebrospinal fluid, urine supernatant, nasopharyngeal swab transport medium (VTM, UTM), bronchoalveolar lavage fluid, cell culture supernatant |
| Performance Range / Specifications | Input volume: 140-560 µL sample; viral nucleic acid recovery: ≥75% at 10³ copies/mL; elution volume: 30-60 µL; A260/A280: 1.6-1.9; column binding capacity: 50 µg total nucleic acid; internal control DNA and RNA compatible |
| Sensitivity / LOD | Limit of detection: 50-100 copies/mL for DNA viruses and 100-200 copies/mL for RNA viruses from 200 µL plasma or serum; carrier RNA enhances recovery of targets below 100 copies/reaction |
| Specificity | Simultaneously recovers DNA and RNA viral genomes; minimal host nucleic acid carryover (typically <5% of total); no cross-contamination between samples with proper technique; validated with diverse viral targets including enveloped, non-enveloped, DNA, and RNA viruses |
| Reaction Conditions / Protocol | Pipette 140-560 µL sample into lysis buffer with carrier RNA; incubate at room temperature for 10 minutes; add ethanol; transfer to spin column; centrifuge at 6,000g for 1 minute; wash with wash buffer AW1; centrifuge; wash with wash buffer AW2; centrifuge; dry spin at full speed for 2 minutes; elute with 30-60 µL elution buffer; incubate 1 minute; centrifuge at 6,000g for 1 minute; total protocol: 25-40 minutes for 1-24 samples |
| Components / Formulation | Lysis buffer (guanidine isothiocyanate, Tris-HCl, EDTA, Triton X-100, poly-A carrier RNA), wash buffer AW1 concentrate (guanidine hydrochloride, Tris-HCl, NaCl, ethanol), wash buffer AW2 concentrate (Tris-HCl, NaCl, ethanol), elution buffer (10 mM Tris-HCl pH 8.0, 0.5 mM EDTA), silica membrane spin columns with collection tubes, loading buffer (ethanol-based) |
| Storage Conditions | All buffers and columns at room temperature (15-25°C); keep lysis buffer bottle tightly closed; protect from direct sunlight; avoid repeated heating and cooling |
| Shelf Life | 18 months from date of manufacture; buffers stable for 6 months after opening when stored at recommended temperature |
| Package Specifications | 50 preparations and 250 preparations; includes spin columns, collection tubes, and all required buffers |
| Product Form | Silica membrane spin columns; liquid buffers; carrier RNA pre-formulated in lysis buffer |
| Quality Control | Each lot tested with quantified viral standards: HBV DNA and HCV RNA recovery ≥75% at 10³ IU/mL; eluate free of PCR inhibitors; column loading capacity verified; no detectable DNase or RNase activity in eluate; inter-column consistency CV <10% |
| Key Features | Simultaneous DNA and RNA recovery; carrier RNA integrated into lysis buffer; spin column format requires only standard microcentrifuge; validated for diverse viral pathogens; ready-to-use buffers; minimal hands-on time |
| Purity | A260/A280: 1.6-1.9 for mixed viral nucleic acid extracts; A260/A230: >1.5 |
| Concentration | Elution volume 30-60 µL; nucleic acid concentration dependent on viral load; typical concentration range 1-30 ng/µL from clinical samples with moderate viremia |
| Activity / Unit Definition | Not applicable for extraction kits |
| Molecular Weight | Not applicable for heterogeneous viral nucleic acid extracts |
| Source / Origin | Silica membrane manufactured from high-purity glass microfiber; carrier RNA is synthetic poly-A RNA; all buffers are molecular biology grade, nuclease-free |
| pH Range / Optimal pH | Lysis buffer pH 6.0-6.5; wash buffer AW1 pH 6.5-7.0; wash buffer AW2 pH 7.0-7.5; elution buffer pH 7.8-8.2 |
| Shipping Conditions | Ambient temperature for all components; no special handling required; stable in transit for up to 3 weeks at temperatures up to 40°C |
| Expiration Date / Stability | 18 months shelf life; real-time stability confirmed by quarterly testing; accelerated stability at 45°C for 21 days shows maintained performance; post-opening stability determined by prevention of buffer contamination |
| Regulatory / Compliance | For research use only; not intended for diagnostic use; manufactured under ISO 9001 quality management system; suitable for epidemiological and surveillance research applications |
| Compatibility | Eluted nucleic acids compatible with all major qPCR and RT-qPCR platforms, digital PCR systems, and isothermal amplification chemistries; spin columns fit standard microcentrifuge rotors for 1.5-2.0 mL tubes |
| Recommended Buffer System | Guanidine isothiocyanate lysis system with integrated carrier RNA; guanidine hydrochloride-containing wash system; Tris-EDTA elution buffer |
| Application Notes / Precautions | Add sample to lysis buffer promptly after preparation; do not allow spin column membrane to dry out between steps; ensure ethanol is added to wash buffers before first use if supplied as concentrates; for maximum recovery from low-titer samples, reduce elution volume to 20-30 µL; use low-retention tubes for eluate storage; avoid repeated freeze-thaw of samples before extraction |
| Batch-to-Batch Consistency | Between-lot viral recovery CV <15%; column binding capacity tested per lot by calf thymus DNA loading; buffer pH and conductivity controlled per batch; carrier RNA integrity and concentration verified per lot preparation |
For research use only, not for clinical use.
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