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| Product Name | Column-Based Genomic DNA Extraction Kit (Whole Blood) |
| Catalog No. | DREK-0011 |
| Description | A silica membrane spin column kit for the rapid purification of high-molecular-weight genomic DNA from whole blood samples. The optimized buffer system contains guanidine hydrochloride and proteinase K for efficient protein digestion and DNA release from leukocytes, followed by selective DNA binding to the silica membrane under high-salt conditions. This classical spin-column method produces DNA with consistent molecular weight and purity suitable for demanding downstream applications. |
| Intended Use | Isolation of genomic DNA from fresh or frozen human whole blood (anticoagulated with EDTA, citrate, or heparin) for PCR, qPCR, Southern blotting, genotyping, NGS library preparation, and long-range PCR applications. |
| Principle / Technology | Whole blood is lysed with a guanidine-containing buffer and proteinase K, releasing genomic DNA from nucleated cells while digesting contaminating proteins. After addition of ethanol, the lysate is centrifuged through a silica membrane spin column where DNA binds. Contaminants are removed by two sequential wash steps, and purified DNA is eluted in low-salt buffer or water. |
| Detection Method | UV spectrophotometry (A260/A280 and A260/A230); agarose gel electrophoresis for size integrity; fluorometric quantitation with PicoGreen or Qubit; PCR amplification of GAPDH and β-globin targets; restriction enzyme digestion |
| Sample Type | Fresh or frozen human whole blood (EDTA, citrate, or heparin anticoagulated); buffy coat; blood stored at 2-8°C for up to 7 days or frozen at -20°C or -80°C |
| Performance Range / Specifications | Input volume: 100-400 µL whole blood; yield from 200 µL normal human blood: 4-12 µg; A260/A280: 1.75-1.90; A260/A230: >1.7; column DNA binding capacity: 30 µg; fragment size: 20-50 kb; DNA suitable for amplification of targets up to 15 kb |
| Sensitivity / LOD | Efficient DNA recovery from as little as 50 µL blood; consistently recovers DNA from leukopenic samples with white blood cell counts as low as 1.0 × 10³/µL; minimum detectable DNA: 1 ng by fluorometric quantitation |
| Specificity | Purified DNA is of human genomic origin; free of co-purified RNA; heme contamination below spectrophotometric detection (A410 <0.01); no detectable bacterial or fungal DNA contamination; suitable for species-specific PCR assays |
| Reaction Conditions / Protocol | Add 20 µL proteinase K to microcentrifuge tube; add 100-400 µL whole blood; adjust volume to 220 µL with PBS if sample <220 µL; add 200 µL lysis/binding buffer; vortex thoroughly; incubate at 56°C for 10 minutes; add 200 µL ethanol; vortex; transfer to spin column; centrifuge at 8,000g for 1 minute; wash with 500 µL wash buffer AW1; centrifuge 1 minute; wash with 500 µL wash buffer AW2; centrifuge 3 minutes at full speed; elute with 50-200 µL elution buffer; incubate 5 minutes at room temperature; centrifuge 1 minute; total protocol: 30-40 minutes |
| Components / Formulation | Proteinase K solution (20 mg/mL), lysis/binding buffer (guanidine hydrochloride, Tris-HCl, EDTA), wash buffer AW1 concentrate (guanidine hydrochloride, Tris-HCl, ethanol), wash buffer AW2 concentrate (Tris-HCl, NaCl, EDTA, ethanol), elution buffer (10 mM Tris-HCl pH 8.5, 0.1 mM EDTA), silica membrane spin columns with collection tubes |
| Storage Conditions | Proteinase K at 2-8°C; all buffers at room temperature (15-25°C); spin columns at room temperature in sealed pouch; protect buffers from direct sunlight |
| Shelf Life | 24 months from date of manufacture; proteinase K stable for 12 months at 2-8°C after opening; buffers stable for 12 months after opening |
| Package Specifications | 50 preparations and 250 preparations (200 µL blood per prep); includes columns and all reagents; proteinase K supplied separately or integrated |
| Product Form | Silica membrane spin columns with collection tubes; liquid buffers; proteinase K solution |
| Quality Control | Each lot validated with pooled human blood: yield 4-12 µg from 200 µL; A260/A280: 1.75-1.90; agarose gel shows DNA >20 kb; GAPDH PCR (1 kb) positive; restriction digestion complete; endotoxin <0.1 EU/µg DNA; DNase/RNase free; lot-to-lot DNA yield CV <15% |
| Key Features | Proven silica membrane technology; consistent DNA yields across donor variability; minimal hands-on time with ready-to-use reagents; broad fragment size recovery; standardized protocol compatible with vacuum manifold processing; cost-effective for high-throughput laboratories |
| Purity | A260/A280: 1.75-1.90; A260/A230: >1.7; protein contamination <0.5%; heme A410 <0.01 |
| Concentration | Elution volume 50-200 µL; typical DNA concentration 40-100 ng/µL from 200 µL normal blood; adjustable elution volume for desired concentration |
| Activity / Unit Definition | Not applicable for DNA extraction kits |
| Molecular Weight | Not applicable for genomic DNA which is heterogeneous in molecular weight |
| Source / Origin | Silica membrane from high-purity borosilicate glass microfiber; proteinase K from recombinant expression in Pichia pastoris; all buffer components are synthetic, molecular biology grade |
| 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 8.3-8.7 |
| Shipping Conditions | Ambient temperature for all components; proteinase K with cold packs for extended transit; no dangerous goods classification |
| Expiration Date / Stability | 24 months shelf life; validated by real-time stability testing at 0, 6, 12, 18, and 24 months; accelerated stability at 37°C for 28 days supports claimed shelf life; column integrity maintained throughout shelf life |
| Regulatory / Compliance | For research use only; not for diagnostic procedures; manufactured under ISO 9001 certified quality management system; Nuclease-free certification per lot |
| Compatibility | Purified DNA suitable for all major PCR, qPCR, and NGS platforms; compatible with vacuum manifolds for high-throughput processing; elution buffer compatible with all common enzymatic reactions |
| Recommended Buffer System | Guanidine hydrochloride-based lysis and binding system; Tris-buffered wash solutions; Tris-EDTA elution for long-term DNA stability |
| Application Notes / Precautions | Pre-warm elution buffer to 56°C for maximum DNA yield; for heparinized blood, add additional proteinase K incubation time; do not allow column membrane to dry completely during wash steps; avoid vigorous pipetting of eluted DNA; for blood stored >7 days, consider fresh proteinase K addition; remove column caps when using vacuum manifold |
| Batch-to-Batch Consistency | Between-lot DNA yield CV <12%; A260/A280 variation <0.04 between lots; column binding capacity verified per lot by λ DNA loading test; buffer conductivity and pH within ±5% specification range per batch; proteinase K specific activity >30 U/mg per batch |
For research use only, not for clinical use.
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