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| Product Name | Column-Based Plant Genomic DNA Extraction Kit |
| Catalog No. | DREK-0019 |
| Description | A silica membrane spin column kit designed for high-quality genomic DNA extraction from diverse plant species. The proprietary CTAB-PVP extraction buffer effectively binds polyphenolic compounds and removes polysaccharides during the chloroform extraction step. The subsequent silica membrane purification eliminates residual contaminants, yielding DNA that meets the purity requirements of demanding applications including next-generation sequencing and genotyping-by-sequencing. |
| Intended Use | Isolation of plant genomic DNA from fresh, frozen, or silica-dried leaf tissue for molecular marker analysis, plant genotyping, GMO detection, phylogenetic studies, DNA barcoding, and plant genomics research. |
| Principle / Technology | Plant tissue is pulverized in liquid nitrogen and lysed in a CTAB buffer containing PVP and β-mercaptoethanol at 65°C. The CTAB detergent solubilizes cellular membranes while PVP sequesters phenolic compounds. A chloroform:isoamyl alcohol extraction removes polysaccharides and proteins. DNA is precipitated from the aqueous phase, redissolved, and further purified through a silica membrane spin column using chaotropic binding chemistry. After washing, purified DNA is eluted in low-salt buffer. |
| Detection Method | UV spectrophotometry at 260/280 nm; agarose gel electrophoresis for integrity; fluorometric quantitation; PCR amplification of plant barcode markers; restriction enzyme digestion; qPCR for GMO and transgene detection |
| Sample Type | Fresh, frozen (-80°C), or silica-gel dried young and mature leaves from model species (Arabidopsis thaliana, Oryza sativa, Nicotiana benthamiana) and major crops (Zea mays, Glycine max, Triticum aestivum, Solanum lycopersicum, Gossypium hirsutum, Brassica napus); seedlings; root tissue |
| Performance Range / Specifications | Input: 100 mg fresh leaf or 30 mg dried tissue; yield: 5-30 µg from 100 mg young leaf; A260/A280: 1.75-1.90; A260/A230: >1.6; fragment size: 20-40 kb; PCR amplification of 2 kb nuclear targets positive under standard conditions |
| Sensitivity / LOD | Effective DNA recovery from 5 mg starting material; single-copy gene detection by PCR in diploid plant genomes; GMO detection at 0.1% contamination level by qPCR |
| Specificity | DNA free of polysaccharides (A260/A230 >1.6) and polyphenols; chloroplast and mitochondrial DNA present at expected genomic proportions; no detectable microbial DNA enrichment; no PCR inhibitors in amplification reactions; species identity confirmed by DNA barcoding markers |
| Reaction Conditions / Protocol | Grind 100 mg tissue to fine powder under liquid nitrogen; add to pre-warmed CTAB buffer with β-mercaptoethanol; incubate at 65°C for 30-60 minutes; add equal volume chloroform:isoamyl alcohol; vortex vigorously; centrifuge 12,000g for 10 minutes; transfer aqueous phase; add 0.7 volumes isopropanol; precipitate at -20°C for 30 minutes; centrifuge; wash pellet with 70% ethanol; air-dry briefly; dissolve in buffer; add binding buffer and ethanol; load onto spin column; centrifuge; wash; elute in 50-100 µL elution buffer; total protocol: 3-4 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, 0.2% β-mercaptoethanol), chloroform:isoamyl alcohol 24:1 (user-supplied), isopropanol (user-supplied), binding buffer (guanidine hydrochloride, Tris-HCl, sodium acetate), wash buffer 1 concentrate, wash buffer 2 concentrate, elution buffer (10 mM Tris-HCl pH 8.5, 0.1 mM EDTA), silica membrane spin columns with collection tubes |
| Storage Conditions | CTAB buffer at room temperature (warm to 65°C before use to dissolve precipitate); wash buffers at room temperature; spin columns at room temperature; protect CTAB buffer from direct light; β-mercaptoethanol at 2-8°C |
| Shelf Life | 24 months for buffers and columns; CTAB buffer stable 12 months at room temperature; β-mercaptoethanol replaced 6 months after opening |
| Package Specifications | 50 and 200 preparation kits; all kit-specific buffers and columns included; user supplies chloroform, isopropanol, ethanol, liquid nitrogen |
| Product Form | Silica membrane spin columns with collection tubes; liquid buffers including CTAB extraction buffer; β-mercaptoethanol vial |
| Quality Control | Each lot tested on Arabidopsis (Columbia-0) and rice (Nipponbare) leaf tissue: yield ≥5 µg from 100 mg; A260/A280: 1.75-1.90; A260/A230: >1.6; PCR for rbcL and ITS markers confirmed; restriction digest complete; no PCR inhibition at 1:10 dilution; species authenticity verified |
| Key Features | CTAB-PVP system effectively removes plant polysaccharides and polyphenols; silica membrane column provides additional purification for purity-sensitive applications; compatible with diverse species across monocots and dicots; validated for GMO detection workflows; reproducible yields across tissue types and developmental stages |
| Purity | A260/A280: 1.75-1.90; A260/A230: >1.6; PCR-verified amplifiability |
| Concentration | Elution volume 50-100 µL; concentration 50-300 ng/µL depending on species and tissue type |
| Activity / Unit Definition | Not applicable for extraction kits |
| Molecular Weight | Not applicable for plant genomic DNA of heterogeneous size |
| Source / Origin | Silica membrane from high-purity quartz microfiber; CTAB from plant-derived source; PVP-40 is synthetic polymer; all buffer components are molecular biology grade, nuclease-free |
| 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 | Ambient temperature for all components; β-mercaptoethanol may require dangerous goods documentation; standard courier acceptable |
| Expiration Date / Stability | 24 months for columns and buffers; CTAB buffer real-time stability confirmed at 18 months; accelerated aging at 37°C supports claimed shelf life; β-mercaptoethanol replaced 6 months after opening |
| Regulatory / Compliance | For research use only; suitable for agricultural biotechnology and plant breeding research; not for diagnostic applications; Nagoya Protocol compliance is the user's responsibility |
| Compatibility | Purified DNA suitable for PCR, qPCR, Sanger sequencing, NGS (whole genome, GBS, RAD-seq), restriction digestion, Southern blotting; compatible with vacuum manifold processing |
| Recommended Buffer System | CTAB-PVP extraction chemistry; guanidine hydrochloride silica membrane binding; Tris-EDTA elution buffer |
| Application Notes / Precautions | Freeze tissue immediately after harvest and store at -80°C; pre-warm CTAB buffer to 65°C; for polyphenol-rich species, increase PVP to 3-4%; avoid DNA shearing by using wide-bore tips; residual ethanol must be completely removed before elution; for GMO detection, run extraction controls alongside samples |
| Batch-to-Batch Consistency | Yield CV <18% between lots on standardized Arabidopsis tissue powder; A260/A280 variation <0.05; column binding capacity >30 µg DNA verified per lot; CTAB buffer pH and conductivity controlled per batch |
For research use only, not for clinical use.
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