- Home
- IVD
- By Technology Types
- By Diseases Types
- By Product Types
- Research
- Resource
- Distributors
- Company
| Product Name | Magnetic Bead-Based Total RNA Extraction Kit |
| Catalog No. | DREK-0006 |
| Description | A magnetic bead-based total RNA purification kit employing a guanidine isothiocyanate-based lysis buffer with β-mercaptoethanol for immediate RNase inactivation. The kit incorporates on-bead DNase I digestion to remove genomic DNA contamination, producing high-integrity total RNA including mRNA, rRNA, tRNA, miRNA, and other small non-coding RNA species. The optimized binding conditions ensure efficient capture of RNA species across a broad size range, while the magnetic bead format eliminates column clogging issues common with viscous tissue lysates. |
| Intended Use | Extraction of total RNA from cultured mammalian cells, animal tissues, bacteria, yeast, and biological fluids for gene expression analysis by RT-qPCR, RT-PCR, Northern blotting, RNA sequencing, and microarray hybridization. |
| Principle / Technology | Samples are homogenized in a guanidine isothiocyanate-based lysis buffer containing β-mercaptoethanol, which simultaneously denatures proteins including RNases while maintaining RNA integrity. Ethanol is added to create optimal binding conditions, and RNA selectively binds to silica-coated magnetic beads. Genomic DNA is removed by on-bead DNase I digestion. Following wash steps, purified total RNA is eluted in RNase-free water or TE buffer. |
| Detection Method | UV spectrophotometry (A260/A280 for purity, A260/A230 for salt contamination); agarose gel electrophoresis with formaldehyde or glyoxal denaturation; Bioanalyzer or TapeStation for RNA Integrity Number (RIN) determination; NanoDrop or Qubit fluorometric quantitation; RT-qPCR of reference genes (GAPDH, ACTB) for functional integrity |
| Sample Type | Cultured mammalian cells (adherent and suspension, fresh or frozen pellets), fresh animal tissues (liver, kidney, brain, spleen, heart, muscle), tissues stored in RNAlater, bacterial and yeast pellets, whole blood leukocyte fraction, and biological fluids (synovial fluid, BAL fluid) |
| Performance Range / Specifications | Binding capacity: 50 µg total RNA per 100 µL bead slurry; input: up to 10^7 cells or 30 mg tissue; yield from 10^6 HeLa cells: 10-30 µg, from 10 mg mouse liver: 30-80 µg; A260/A280: 1.9-2.1; A260/A230: >1.8; RIN >7.0 for properly handled tissues; RNA includes miRNA fraction (validated by small RNA Bioanalyzer trace) |
| Sensitivity / LOD | Recovers total RNA from as few as 100 cells using scaled-down protocol; detects low-abundance transcripts at <5 copies per cell by RT-qPCR; miRNA recovery efficiency >80% compared to dedicated miRNA isolation methods |
| Specificity | Recovers total RNA including mRNA, rRNA, tRNA, miRNA, piRNA, and other small non-coding RNAs without size fractionation; on-bead DNase I digestion eliminates genomic DNA contamination (validated by minus-RT PCR); no detectable RNase activity in eluted RNA stored at -80°C for 12 months |
| Reaction Conditions / Protocol | Homogenize sample in 350-600 µL lysis buffer with β-mercaptoethanol (added fresh); for tissue, use rotor-stator homogenizer or bead mill; add equal volume 70% ethanol; add magnetic bead suspension; incubate 5 minutes at room temperature; separate beads; wash with wash buffer 1; apply DNase I mixture directly to beads; incubate 15 minutes at room temperature; wash with wash buffer 1; wash twice with wash buffer 2; dry beads; elute in 30-100 µL RNase-free water; total time: 45-60 minutes including DNase digestion |
| Components / Formulation | Lysis buffer (guanidine isothiocyanate, sodium citrate, N-lauroylsarcosine, β-mercaptoethanol), wash buffer 1 (guanidine hydrochloride, Tris-HCl, ethanol), wash buffer 2 concentrate (Tris-HCl, NaCl, ethanol), magnetic silica bead suspension (50 mg/mL), DNase I (lyophilized, RNase-free), DNase reconstitution buffer (Tris-HCl, MgCl2, CaCl2), RNase-free water, magnetic separation rack; β-mercaptoethanol required but provided separately |
| Storage Conditions | Lysis buffer at room temperature (15-25°C) protected from light; DNase I lyophilized at -20°C, reconstituted at 2-8°C for up to 4 weeks; wash buffers at room temperature; magnetic bead suspension at 2-8°C; all components RNase-free |
| Shelf Life | 24 months for buffers and magnetic beads; lyophilized DNase I 24 months at -20°C; reconstituted DNase I 4 weeks at 2-8°C |
| Package Specifications | 50 preparations and 200 preparations; includes DNase I and all required buffers; magnetic rack sold separately |
| Product Form | Liquid buffers; silica-coated magnetic bead suspension; lyophilized DNase I with reconstitution buffer; β-mercaptoethanol vial |
| Quality Control | Each lot validated with HeLa cell RNA: yield ≥15 µg/10^6 cells; A260/A280: 1.9-2.1; RIN ≥8.0; gDNA removal confirmed by minus-RT qPCR (no amplification); miRNA recovery verified by miR-16 and miR-21 RT-qPCR; RNase-free certification by incubation with fluorescent RNA substrate; functional RT-qPCR efficiency for six reference genes: 90-110% |
| Key Features | On-bead DNase digestion eliminates separate DNA removal step; magnetic bead format prevents column clogging; broad RNA size recovery including miRNA; rapid 45-minute protocol; no phenol-chloroform extraction; compatible with fresh, frozen, and RNAlater samples |
| Purity | A260/A280: 1.9-2.1; A260/A230: >1.8; no detectable protein by Bradford assay; gDNA contamination below qPCR detection limit |
| Concentration | Elution volume 30-100 µL; RNA concentration 100-500 ng/µL for cell samples, 200-800 ng/µL for tissue samples; total yield dependent on sample type and cell count |
| Activity / Unit Definition | Not applicable for RNA extraction kits |
| Molecular Weight | Not applicable; total RNA is a heterogeneous mixture of species ranging from ~20 nucleotides (miRNA) to >10 kb (mRNA) |
| Source / Origin | Silica-coated magnetic beads synthesized via sol-gel method; DNase I from bovine pancreas, chromatography purified; all buffers use molecular biology grade reagents; no animal-derived RNase inhibitors used in buffer formulation |
| pH Range / Optimal pH | Lysis buffer pH 6.0-6.5; wash buffer 1 pH 6.5-7.0; wash buffer 2 pH 7.0-7.5; DNase buffer pH 7.6-7.8; elution water pH 6.5-7.5 |
| Shipping Conditions | Ambient temperature for buffers and beads; DNase I shipped on dry ice; β-mercaptoethanol requires dangerous goods declaration for air transport |
| Expiration Date / Stability | 24 months from manufacture; accelerated stability at 37°C for 14 days confirms RNA integrity preservation; DNase I retains >85% specific activity after 4 weeks at 2-8°C; lysis buffer DTT equivalent (β-mercaptoethanol) antioxidant capacity verified stable for 18 months in sealed container |
| Regulatory / Compliance | For research use only; not intended for diagnostic procedures; RNase-free certification provided per lot; manufactured in ISO 9001 certified facility |
| Compatibility | Purified RNA suitable for RT-qPCR, RT-PCR, cDNA synthesis, Northern blotting, microarray, RNA-seq (including small RNA-seq), and RACE; compatible with magnetic particle processors; recommended for use with RNase-free labware and dedicated RNA workspace |
| Recommended Buffer System | Guanidine isothiocyanate-sodium citrate lysis; Tris-buffered ethanol washes; DNase I digestion in Tris-MgCl2-CaCl2 buffer; nuclease-free water elution |
| Application Notes / Precautions | Maintain an RNase-free work environment throughout processing; add β-mercaptoethanol to lysis buffer immediately before use; for tissue samples, ensure complete homogenization to maximize yield; avoid over-drying magnetic beads after final wash; store RNA at -80°C for long-term storage or -20°C for short-term; use RNase-free pipette tips and microcentrifuge tubes; for bacterial samples, pretreatment with lysozyme may improve lysis of Gram-positive species |
| Batch-to-Batch Consistency | Between-lot CV for total RNA yield <12%; RIN variation between lots <0.5 units for standardized HeLa lysate; DNase I activity verified per batch ensures complete gDNA removal; buffer pH and conductivity controlled to ±0.1 pH and ±5% conductivity |
For research use only, not for clinical use.
|
There is no product in your cart. |
Copyright © 2026 Alta DiagnoTech. All rights reserved.
