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| Product Name | Magnetic Bead-Based Genomic DNA Extraction Kit (Plant) |
| Catalog No. | DREK-0004 |
| Description | A magnetic bead-based genomic DNA extraction kit specifically formulated for plant tissues. The kit addresses common challenges in plant DNA isolation including high polysaccharide and polyphenol content, rigid cell walls, and the presence of secondary metabolites that can inhibit downstream enzymatic reactions. A specialized CTAB-containing lysis buffer combined with PVP and β-mercaptoethanol effectively removes phenolic compounds and polysaccharides, while the magnetic bead-based purification ensures consistent DNA quality across diverse plant species. |
| Intended Use | Isolation of genomic DNA from fresh, frozen, or silica-dried plant tissues including leaves, roots, seeds, fruits, and seedlings for plant genotyping, GMO detection, marker-assisted breeding, phylogenetic analysis, and plant-pathogen interaction studies. |
| Principle / Technology | Plant tissue is ground to a fine powder in liquid nitrogen and homogenized in a pre-warmed CTAB-based extraction buffer containing PVP and β-mercaptoethanol. After chloroform extraction to remove polysaccharides and proteins, the aqueous phase is mixed with binding buffer and silica-coated magnetic beads. DNA binds to beads under high-salt conditions. Following magnetic capture and ethanol-based washes, purified plant genomic DNA is eluted in low-salt buffer. |
| Detection Method | UV spectrophotometry (A260/A280, A260/A230); agarose gel electrophoresis; fluorometric quantitation; PCR amplification of plant-specific loci (rbcL, ITS, matK); restriction enzyme digestion for quality confirmation |
| Sample Type | Fresh, frozen (at -80°C), or silica gel-dried plant leaves (young, mature, and senescent), roots, stems, seeds, fruits, and seedlings; tested with Arabidopsis thaliana, Oryza sativa (rice), Zea mays (maize), Glycine max (soybean), Nicotiana tabacum (tobacco), Triticum aestivum (wheat), and Populus species (poplar) |
| Performance Range / Specifications | Input: 50-100 mg fresh leaf tissue or 20-50 mg dried tissue; yield: 5-25 µg from 100 mg young Arabidopsis leaf, 3-15 µg from 100 mg rice leaf; A260/A280: 1.75-1.90; A260/A230: >1.6; fragment size: 20-40 kb; DNA suitable for amplification of targets up to 5 kb |
| Sensitivity / LOD | Reliably recovers plant genomic DNA from 10 mg starting material; effective with high-polysaccharide species including succulents and cacti; detects single-copy nuclear genes by PCR in Arabidopsis and rice genomes |
| Specificity | DNA free of co-purified polysaccharides and polyphenols (indicated by A260/A230 >1.6); no detectable RNase activity; minimal chloroplast and mitochondrial DNA enrichment relative to nuclear genome representation; no PCR inhibitors in standard amplification reactions |
| Reaction Conditions / Protocol | Grind 50-100 mg plant tissue to fine powder under liquid nitrogen; transfer to pre-warmed CTAB extraction buffer (65°C) containing PVP and β-mercaptoethanol; incubate at 65°C for 30-60 minutes with intermittent mixing; add equal volume chloroform:isoamyl alcohol (24:1), vortex, centrifuge 12,000g for 10 minutes; transfer aqueous phase; add 0.7 volumes isopropanol and binding buffer; add magnetic bead suspension; incubate 10 minutes at room temperature; separate beads; wash once with wash buffer 1 and twice with wash buffer 2; air-dry; elute in 50-100 µL Tris buffer at 65°C; total protocol: approximately 2-3 hours |
| Components / Formulation | CTAB extraction buffer (2% CTAB, 100 mM Tris-HCl pH 8.0, 20 mM EDTA, 1.4 M NaCl, 2% PVP-40), β-mercaptoethanol (added fresh), binding buffer (guanidine hydrochloride, Tris-HCl, isopropanol), magnetic silica bead suspension (50 mg/mL), wash buffer 1 concentrate (guanidine hydrochloride, Tris-HCl, ethanol), wash buffer 2 concentrate (Tris-HCl, ethanol), elution buffer (10 mM Tris-HCl pH 8.5), magnetic separation rack; chloroform:isoamyl alcohol (24:1) required but not supplied |
| Storage Conditions | CTAB extraction buffer at room temperature or 37°C to prevent precipitation; β-mercaptoethanol at 2-8°C; magnetic bead suspension at 2-8°C; wash buffers at room temperature; all buffers protected from light |
| Shelf Life | 24 months for dry components; CTAB extraction buffer stable 12 months at room temperature; magnetic beads 18 months at 2-8°C |
| Package Specifications | 50 preparations and 200 preparations; all reagents included except chloroform:isoamyl alcohol; magnetic rack available separately |
| Product Form | Liquid buffers; silica-coated magnetic bead suspension; CTAB powder for buffer preparation |
| Quality Control | Each lot tested with Arabidopsis (Columbia-0) leaf: yield ≥5 µg from 100 mg; A260/A280: 1.75-1.90; A260/A230: >1.6; PCR amplification of rbcL gene (1.3 kb) confirmed; no qPCR inhibition at 1:10 dilution; tested across rice, maize, tomato, and tobacco for species diversity validation |
| Key Features | CTAB-PVP system effectively removes plant polysaccharides; magnetic bead format eliminates columns that clog with viscous samples; compatible with diverse plant species including polyphenol-rich and mucilaginous plants; no phenol extractions; high-purity DNA for sensitive genomic applications |
| Purity | A260/A280: 1.75-1.90; A260/A230: >1.6 (indicating low polysaccharide/polyphenol carryover) |
| Concentration | Elution in 50-100 µL; DNA concentration typically 50-300 ng/µL dependent on species and tissue type |
| Activity / Unit Definition | Not applicable for extraction kits |
| Molecular Weight | Not applicable for heterogeneous genomic DNA from plant sources |
| Source / Origin | Silica-coated magnetic beads manufactured via Stöber process with surface functionalization; CTAB from plant-derived source; all buffer components are synthetic, molecular biology grade |
| pH Range / Optimal pH | CTAB extraction 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 | All components ship at ambient temperature except β-mercaptoethanol which ships as dangerous goods under IATA regulations; magnetic beads shipped with cold packs for extended transit |
| Expiration Date / Stability | 24 months when stored as directed; real-time stability data available at 24 months; CTAB buffer visually inspected for precipitation before use; β-mercaptoethanol replaced every 6 months after opening |
| Regulatory / Compliance | For research use only; not for diagnostic applications; suitable for agricultural biotechnology research; complies with Nagoya Protocol guidelines for genetic resource utilization record-keeping |
| Compatibility | Purified DNA suitable for PCR, qPCR, genotyping-by-sequencing (GBS), restriction-site associated DNA sequencing (RAD-seq), whole-genome sequencing, and marker-assisted selection workflows; compatible with magnetic particle handler platforms |
| Recommended Buffer System | CTAB-based extraction with PVP and β-mercaptoethanol as reducing and complexing agents; guanidine hydrochloride binding system; Tris-EDTA elution buffer |
| Application Notes / Precautions | Pre-warm CTAB buffer to 65°C before use to dissolve any precipitate; add β-mercaptoethanol to CTAB buffer just before use; for seeds, soak in water overnight at 4°C before processing; for mature leaves with high secondary metabolites, increase PVP and β-mercaptoethanol concentrations; avoid mechanical shearing during mixing steps; use wide-bore pipette tips for transferring eluted DNA |
| Batch-to-Batch Consistency | Between-lot DNA yield CV <18% across multiple plant species; A260/A280 variation <0.05 between lots; binding capacity verified at ≥15 µg DNA per preparation; CTAB buffer lot pre-tested for extraction efficiency on standardized Arabidopsis tissue powder |
For research use only, not for clinical use.
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