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| Product Name | Automated 96-Well Plate RNA Extraction Kit |
| Catalog No. | DREK-0021 |
| Description | A high-throughput magnetic bead-based total RNA extraction kit designed for fully automated 96-well processing on open-deck liquid handling systems. The kit provides pre-formulated, automation-ready reagents including a guanidine-based lysis buffer with integrated β-mercaptoethanol replacement, magnetic silica beads in a dispensing-compatible suspension, and an on-deck DNase I treatment step for genomic DNA removal. The workflow delivers consistent RNA integrity and yield across all 96 wells, supporting large-scale gene expression studies and transcriptomic projects. |
| Intended Use | Automated high-throughput extraction of total RNA from cultured cells, tissue homogenates, and blood leukocyte fractions in 96-well plate format for large-scale gene expression analysis, biomarker discovery, and transcriptomic studies. |
| Principle / Technology | Samples in a 96-well plate are lysed with guanidine isothiocyanate-based buffer containing a non-thiol reducing agent. The liquid handler adds magnetic bead suspension after ethanol addition, and RNA binds to silica-coated beads. The platform performs magnetic bead capture, supernatant removal, and serial ethanol-based washes. A DNase I solution is dispensed onto the beads for on-bead genomic DNA digestion, followed by additional washes. Purified total RNA is eluted in RNase-free water and transferred to an output plate. |
| Detection Method | UV spectrophotometry via plate reader; fluorometric RNA quantitation (96-well format); Bioanalyzer or TapeStation for RNA integrity from representative wells; RT-qPCR of reference genes for functional quality; RNA-seq library metrics for validation plates |
| Sample Type | Cultured mammalian cell pellets (up to 5 × 10⁶ cells per well), fresh or frozen tissue homogenates (pre-prepared, up to 20 mg tissue per well), white blood cell pellets; samples must be prepared in single-cell suspension or homogeneous lysate prior to loading |
| Performance Range / Specifications | Throughput: 96 samples per run in 75-100 minutes; input: up to 5 × 10⁶ cells or 20 mg tissue; yield: 5-20 µg per well from 10⁶ HeLa cells; A260/A280: 1.9-2.1; well-to-well yield CV: <12%; plate-to-plate CV: <15%; elution volume: 30-100 µL; on-deck DNase treatment eliminates genomic DNA |
| Sensitivity / LOD | RNA recovery from as few as 10³ cells per well using miniaturized protocol; low-abundance mRNA detection at <5 copies per cell by RT-qPCR; miRNA recovery efficiency >70% compared to dedicated miRNA methods; DNase treatment eliminates gDNA to below qPCR detection |
| Specificity | Total RNA recovery including mRNA, rRNA, tRNA, and miRNA; no residual genomic DNA after on-deck DNase treatment; no cross-well contamination; consistent RNA integrity across plate positions; RNase-free certified all components |
| Reaction Conditions / Protocol | Program liquid handler: load sample plate with cell pellets in lysis buffer; incubate 5 minutes room temperature; add ethanol and magnetic bead suspension; mix; incubate 5 minutes; magnetically capture beads; remove supernatant; wash with wash buffer 1; capture; wash with wash buffer 2; capture; add DNase I master mix; incubate 15 minutes at room temperature; add wash buffer 1; capture; wash with wash buffer 2 twice; air-dry beads; add RNase-free water; incubate 5 minutes; capture beads; transfer RNA eluate to output plate; total deck time: 75-100 minutes |
| Components / Formulation | Lysis buffer (guanidine isothiocyanate, Tris-HCl, EDTA, reducing agent) in bulk bottle, magnetic silica bead suspension in bulk dispensing bottle, wash buffer 1 concentrate in bulk, wash buffer 2 concentrate in bulk, DNase I (lyophilized) with reconstitution buffer, RNase-free water, 96-well deep-well processing plates, 96-well elution plates, adhesive seals, magnetic separation plate carrier |
| Storage Conditions | Lysis buffer at room temperature protected from light; DNase I at -20°C lyophilized, reconstituted at 2-8°C for up to 1 week; wash buffers at room temperature; magnetic beads at 2-8°C; pre-filled plates in sealed pouches at room temperature |
| Shelf Life | 18 months for buffers; DNase I 24 months lyophilized at -20°C; magnetic bead suspension 18 months at 2-8°C; reconstituted DNase I 1 week at 2-8°C |
| Package Specifications | 4 × 96 preparation kits and 10 × 96 preparation kits; includes all reagents, plates, seals, and DNase I; compatible with major liquid handler platforms |
| Product Form | Bulk liquid reagents; magnetic bead suspension in dispenser-compatible bottle; pre-filled reagent plates; lyophilized DNase I; consumable plates and seals |
| Quality Control | Each lot validated with HeLa cell reference standard across full 96-well test plate: mean yield ≥10 µg/10⁶ cells; A260/A280: 1.9-2.1; RIN ≥7.5 for all tested wells; gDNA-free confirmed by minus-RT qPCR; intra-plate yield CV <12%; DNase/RNase free |
| Key Features | Walk-away automation with pre-configured reagent kits; on-deck DNase digestion for genomic DNA removal; consistent RNA integrity across all 96 wells; optimized for major open-deck liquid handlers; bulk reagent format for cost efficiency; output plate directly compatible with cDNA synthesis and library preparation |
| Purity | A260/A280: 1.9-2.1; A260/A230: >1.8; genomic DNA contamination below qPCR detection |
| Concentration | Adjustable elution 30-100 µL; concentration 50-500 ng/µL depending on input; concentration CV <12% across plate |
| Activity / Unit Definition | Not applicable for RNA extraction kits |
| Molecular Weight | Not applicable for total RNA of heterogeneous size distribution |
| Source / Origin | Silica-coated magnetic beads from sol-gel synthesis; DNase I from bovine pancreas, RNase-free purified; all buffers molecular biology grade, nuclease-free certified; plates from virgin polypropylene |
| pH Range / Optimal pH | Lysis buffer pH 6.0-6.5; wash buffers pH 6.5-7.5; DNase buffer pH 7.5-7.8; elution water pH 6.5-7.5 |
| Shipping Conditions | Ambient temperature for bulk buffers and plates; DNase I on dry ice; magnetic beads with cold packs; standard logistics for most components |
| Expiration Date / Stability | 18 months for buffers and plates; DNase I lyophilized 24 months at -20°C; magnetic beads 18 months at 2-8°C; real-time stability confirmed at 18 months with RNA integrity verification |
| Regulatory / Compliance | For research use only; not for diagnostic procedures; RNase-free certification per lot; manufactured under ISO 9001 and ISO 13485 quality management |
| Compatibility | Validated on Tecan Freedom EVO, Hamilton Microlab Star, Beckman Coulter Biomek i-Series, and Agilent Bravo platforms; eluted RNA compatible with RT-qPCR, microarray, and RNA-seq library preparation; plate format directly integrates with downstream automation |
| Recommended Buffer System | Guanidine isothiocyanate lysis with non-thiol reducing agent; silica bead binding chemistry; Tris-buffered washes; RNase-free water elution; DNase I in Tris-MgCl2 digestion buffer |
| Application Notes / Precautions | Program sufficient bead drying time after final wash; calibrate magnet engagement time for specific platform; validate DNase incubation time for specific plate type; use plate seals during incubation to prevent evaporation and RNase contamination; pre-cool elution plate if processing heat-sensitive samples; for tissues, ensure complete homogenization before loading onto deck |
| Batch-to-Batch Consistency | Intra-plate yield CV <12%; inter-lot yield CV <15%; DNase I activity verified per batch for complete gDNA removal; magnetic bead binding capacity >50 µg RNA per preparation verified per batch; RNase-free certification per batch |
For research use only, not for clinical use.
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