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| Product Name | Magnetic Bead-Based Genomic DNA Extraction Kit (Tissue) |
| Catalog No. | DREK-0003 |
| Description | A magnetic bead-based kit formulated for the extraction of high-quality genomic DNA from a diverse range of solid tissues including fresh, frozen, and RNAlater-preserved specimens. The enzymatic tissue lysis step incorporates optimized collagenase and proteinase K digestion conditions to ensure complete tissue disruption and release of genomic DNA from fibrous and lipid-rich tissues. The isotropic magnetic bead technology provides rapid binding kinetics and high DNA recovery across a broad input mass range. |
| Intended Use | Extraction of genomic DNA from fresh or frozen animal and human tissue samples including liver, kidney, brain, heart, muscle, skin, spleen, lung, and adipose tissue for molecular biology and genomic applications. |
| Principle / Technology | Tissue samples are mechanically disrupted in a specialized tissue digestion buffer containing SDS and EDTA, followed by incubation with proteinase K at 56°C to digest proteins and release DNA from chromatin. After complete digestion, guanidine hydrochloride binding buffer is added to promote DNA binding to silica-coated magnetic beads. Beads are magnetically captured, washed to remove tissue debris and inhibitors, and purified DNA is eluted in low-ionic-strength buffer. |
| Detection Method | UV spectrophotometry; agarose gel electrophoresis; fluorometric quantitation; PCR amplification of housekeeping genes and mitochondrial targets; restriction enzyme digestion for quality assessment |
| Sample Type | Fresh mammalian and avian tissues (liver, kidney, brain, heart, muscle, spleen, lung, skin, adipose); frozen tissues stored at -20°C or -80°C; RNAlater-preserved tissues (after PBS rinse); formalin-fixed tissues (requires heat-induced antigen retrieval pretreatment, not included); tail snips and ear punches from rodent models |
| Performance Range / Specifications | Input: 10-30 mg solid tissue; yield: 5-30 µg from 25 mg mouse liver, 3-10 µg from 25 mg skeletal muscle, 2-8 µg from 25 mg brain tissue; A260/A280: 1.75-1.90; A260/A230: >1.7; fragment size: 25-50 kb depending on tissue freshness and handling |
| Sensitivity / LOD | Reliably recovers genomic DNA from 5 mg tissue biopsies; picogram-level DNA detectable by qPCR; effective with partially degraded specimens |
| Specificity | High specificity for double-stranded genomic DNA; minimal RNA co-purification (<2% by fluorometric RNA assay); inhibitor-free DNA confirmed by consistent qPCR amplification across sample types; no detectable DNase activity in eluate |
| Reaction Conditions / Protocol | Excise 10-30 mg tissue and mince finely; add tissue digestion buffer and proteinase K; incubate at 56°C until complete digestion (1-3 hours for most tissues, overnight for fibrous tissues); vortex periodically during digestion; add binding buffer and isopropanol; add magnetic bead suspension; incubate 10 minutes at room temperature; magnetically separate beads; wash once with wash buffer 1 and twice with wash buffer 2; air-dry beads; elute in 50-100 µL Tris-EDTA buffer at 56°C for 10 minutes; total time: 1.5-4 hours excluding overnight digestion option |
| Components / Formulation | Tissue digestion buffer (Tris-HCl pH 8.0, EDTA, SDS, NaCl), proteinase K solution (20 mg/mL), binding buffer (guanidine hydrochloride, Tris-HCl, isopropanol), magnetic silica bead suspension (50 mg/mL), wash buffer 1 concentrate (guanidine hydrochloride, Tris-HCl, NaCl, ethanol), wash buffer 2 concentrate (Tris-HCl, NaCl, ethanol), elution buffer (10 mM Tris-HCl pH 8.5, 0.1 mM EDTA), magnetic separation rack |
| Storage Conditions | Tissue digestion buffer and other buffers at room temperature; proteinase K at 2-8°C; magnetic bead suspension at 2-8°C; protect buffers from light; avoid freezing magnetic beads |
| Shelf Life | 24 months for buffers; proteinase K stable 12 months at 2-8°C after first use; magnetic bead suspension stable 18 months at 2-8°C |
| Package Specifications | 50 preparations (up to 30 mg tissue per prep) and 200 preparations; includes all reagents and consumables; magnetic rack sold separately |
| Product Form | Liquid buffers; silica-coated magnetic bead suspension; proteinase K in glycerol-based buffer |
| Quality Control | Each lot validated with mouse liver tissue: yield ≥10 µg from 25 mg; A260/A280: 1.75-1.90; PCR amplification of 5 kb GAPDH target; no qPCR inhibition at 1:10 dilution; tissue panel test includes muscle, brain, and kidney; endotoxin <0.1 EU/µg DNA |
| Key Features | Effective with fibrous and tough tissues; overnight digestion option for difficult samples; no organic solvent extraction required; magnetic bead format enables scalable throughput from single samples to 96-well processing; inhibitor-free DNA suitable for NGS; room temperature stable buffers for convenient storage |
| Purity | A260/A280: 1.75-1.90; A260/A230: >1.7; protein contamination <1% by Bradford assay |
| Concentration | Elution volume 50-100 µL; typical concentration 50-300 ng/µL from 25 mg input depending on tissue type and cellularity |
| Activity / Unit Definition | Not applicable for extraction kits |
| Molecular Weight | Not applicable for heterogeneous genomic DNA |
| Source / Origin | Silica-coated magnetic beads are manufactured through sol-gel synthesis; proteinase K expressed recombinantly; all buffer components are of molecular biology grade with certificates of analysis |
| pH Range / Optimal pH | Tissue digestion buffer pH 7.8-8.2; binding buffer pH 5.5-6.5; wash buffers pH 6.5-7.5; elution buffer pH 8.3-8.7 |
| Shipping Conditions | Ambient temperature for dry reagents; proteinase K shipped with cold packs; magnetic beads shipped at 2-8°C or ambient with cold packs depending on distance |
| Expiration Date / Stability | 24 months shelf life from date of manufacture when stored as recommended; real-time stability verified at 24 months; accelerated degradation studies at 40°C support claimed shelf life |
| Regulatory / Compliance | For research use only; not intended for human or animal diagnostic or therapeutic applications; components manufactured under ISO 9001 quality system |
| Compatibility | Purified DNA suitable for PCR, real-time PCR, digital PCR, Sanger sequencing, NGS library construction, restriction digestion, Southern blotting, and microarray analysis; compatible with magnetic particle processors including KingFisher systems |
| Recommended Buffer System | Tris-SDS tissue digestion buffer; guanidine hydrochloride binding chemistry; Tris-EDTA elution optimized for DNA stability |
| Application Notes / Precautions | For adipose and brain tissues with high lipid content, increase digestion time and consider PBS wash step; remove excess ethanol from wash buffer 2 before elution to avoid carryover; pre-warm elution buffer for maximum yield; if tissue digestion is incomplete after 3 hours, add fresh proteinase K and continue incubation; do not use more than 30 mg tissue per column equivalent to avoid bead saturation |
| Batch-to-Batch Consistency | Yield CV <15% between lots tested on standardized mouse liver homogenate; A260/A280 variation between lots <0.05 units; binding capacity verified per lot: ≥20 µg DNA per 50 µL bead slurry; absence of DNase confirmed by fluorogenic substrate assay per batch |
For research use only, not for clinical use.
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