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| Product Name | Column-Based Genomic DNA Extraction Kit (Tissue) |
| Catalog No. | DREK-0012 |
| Description | A spin column-based genomic DNA purification kit designed for high-yield extraction from animal tissues. The kit combines an SDS-based proteinase K digestion step with silica membrane purification, providing a straightforward workflow that produces DNA of high molecular weight and purity. The optimized lysis buffer formulation accommodates a broad range of tissue types including fibrous, lipid-rich, and tough connective tissues. |
| Intended Use | Isolation of genomic DNA from fresh or frozen animal tissues (rodent, livestock, fish, and other vertebrates) for PCR-based genotyping, quantitative PCR, Southern blot analysis, and targeted sequencing applications. |
| Principle / Technology | Tissue is lysed by incubation with an SDS-containing digestion buffer and proteinase K, which digests structural proteins and releases genomic DNA. After complete digestion, a guanidine hydrochloride-based binding buffer and ethanol are added to adjust conditions for DNA binding to the silica membrane. The lysate is centrifuged through the spin column, washed twice to remove tissue debris and contaminants, and purified DNA is eluted. |
| Detection Method | UV spectrophotometry for purity and concentration; 0.8% agarose gel electrophoresis for molecular weight assessment; fluorometric quantitation for precise yield; restriction enzyme digestion for quality validation; PCR amplification of species-specific loci |
| Sample Type | Fresh and frozen tissues from mouse, rat, rabbit, porcine, bovine, ovine, piscine (fish), and avian sources; tissue types include liver, kidney, heart, lung, muscle, brain, spleen, tail, ear punch, and skin |
| Performance Range / Specifications | Input: 10-25 mg tissue; yield from 25 mg mouse liver: 10-30 µg; from 25 mg mouse tail: 5-15 µg; from 25 mg muscle: 3-10 µg; A260/A280: 1.75-1.90; A260/A230: >1.7; column binding capacity: 40 µg; fragment size: 20-50 kb |
| Sensitivity / LOD | Effective DNA recovery from 5 mg tissue samples; successful PCR amplification from single mouse ear punch lysate; recovers sufficient DNA for >500 PCR reactions from a single 25 mg mouse-tail preparation |
| Specificity | Purified DNA is of animal genomic origin; RNA contamination below agarose gel detection; no detectable protein by Bradford assay; minimal mitochondrial DNA enrichment; no PCR inhibitors detected by internal amplification control |
| Reaction Conditions / Protocol | Excise 10-25 mg tissue, mince into small pieces; add 180 µL tissue digestion buffer and 20 µL proteinase K; vortex; incubate at 56°C until complete lysis (1-3 hours for most tissues); vortex periodically; add 200 µL binding buffer and 200 µL ethanol; vortex; transfer to spin column; centrifuge at 8,000g for 1 minute; wash with 500 µL wash buffer AW1; centrifuge; wash with 500 µL wash buffer AW2; centrifuge 3 minutes at full speed; elute with 50-200 µL elution buffer; incubate 5 minutes; centrifuge; total time: 1.5-4 hours depending on tissue type |
| Components / Formulation | Tissue digestion buffer (Tris-HCl pH 8.0, EDTA, SDS, NaCl), proteinase K solution (20 mg/mL in glycerol buffer), binding buffer (guanidine hydrochloride, Tris-HCl, ethanol), wash buffer AW1 concentrate (guanidine hydrochloride, Tris-HCl, NaCl, 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 | Tissue digestion buffer and other buffers at room temperature; proteinase K at 2-8°C; spin columns at room temperature; protect buffers from light |
| Shelf Life | 24 months for buffers and columns; proteinase K 12 months at 2-8°C after first use |
| Package Specifications | 50 preparations and 250 preparations; all columns, tubes, and reagents included |
| Product Form | Silica membrane spin columns with collection tubes; liquid buffers; proteinase K solution |
| Quality Control | Each lot validated with mouse liver and tail tissue: yield in specified range; A260/A280: 1.75-1.90; amplification of Gapdh gene (1 kb fragment) confirmed; no PCR inhibition at 1:10 dilution; tissue panel tests across liver, muscle, and brain for yield consistency |
| Key Features | Simple spin column workflow; effective with diverse tissue types; SDS-based lysis for complete protein removal; consistent yield across specimen variability; ready-to-use buffers require no preparation; compatible with vacuum manifold processing for throughput |
| Purity | A260/A280: 1.75-1.90; A260/A230: >1.7; protein contamination <0.3% by BCA assay |
| Concentration | Elution volume 50-200 µL; DNA concentration 50-300 ng/µL depending on tissue cellularity; adjustable elution volume to achieve desired concentration |
| Activity / Unit Definition | Not applicable for DNA extraction kits |
| Molecular Weight | Not applicable for genomic DNA of heterogeneous molecular weight distribution |
| Source / Origin | Silica membrane from high-purity quartz microfiber; proteinase K is recombinant, expressed in Pichia pastoris; all buffers use USP/EP-grade chemicals |
| pH Range / Optimal pH | Tissue digestion buffer pH 7.8-8.2; binding buffer pH 5.5-6.5; 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 shipping for all components; proteinase K shipped with cold packs for long-distance transport |
| Expiration Date / Stability | 24 months unopened; annual real-time stability testing confirms performance; proteinase K retains >90% activity after 12 months at 2-8°C post-opening; column binding capacity remains >90% of initial specification through shelf life |
| Regulatory / Compliance | For research use only; suitable for animal research and veterinary research applications; not for human clinical diagnostic use; manufactured under ISO 9001 quality principles |
| Compatibility | Eluted DNA suitable for standard and real-time PCR, Sanger sequencing, NGS library construction, restriction digestion, and Southern blotting; compatible with vacuum manifold systems; elution in water, TE, or EB buffer all acceptable |
| Recommended Buffer System | Tris-SDS tissue digestion buffer; guanidine hydrochloride binding chemistry; ethanol-containing wash buffers; Tris-EDTA elution for DNA stability |
| Application Notes / Precautions | For fatty tissues (brain, adipose), trim excess fat before processing; for fibrous tissues (heart, muscle, skin), extend digestion time and vortex frequently; do not exceed recommended tissue input to avoid column clogging; pre-warm elution buffer for maximum yield; if using vacuum manifold, adjust vacuum pressure to avoid membrane damage; for tail snips from young animals (<14 days), reduce digestion time by half |
| Batch-to-Batch Consistency | Between-lot DNA yield CV <15% on standardized mouse liver homogenate; A260/A280 <0.05 variation between lots; binding capacity ≥40 µg per column verified by λ DNA loading test per lot; DNase/RNase tested per batch; buffer pH tolerance ±0.1 units per batch |
For research use only, not for clinical use.
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