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| Product Name | Oligo-dT Magnetic Beads (mRNA Capture) |
| Catalog No. | MAGBEA-0016 |
| Description | Oligo-dT functionalized superparamagnetic beads for the selective capture and purification of polyadenylated mRNA from total RNA preparations and direct cell or tissue lysates. The 25-mer deoxythymidine oligonucleotides are covalently coupled to the bead surface via a stable spacer arm, enabling high-affinity hybridization to the poly-A tails of eukaryotic mRNA. The magnetic bead format provides rapid mRNA enrichment without columns, spin steps, or organic solvents, making it suitable for both manual and automated transcriptomics workflows. |
| Intended Use | Isolation of poly(A)+ mRNA from total RNA preparations, cell lysates, and tissue homogenates for RNA-seq library preparation, cDNA synthesis, RT-qPCR gene expression analysis, and eukaryotic transcriptomics research. |
| Principle / Technology | Synthetic 5'-amino-modified oligo-dT(25) oligonucleotides are covalently linked to the bead surface through a hydrophilic spacer arm. Under high-salt hybridization conditions, the oligo-dT sequence hybridizes via Watson-Crick base pairing to the 3' poly(A) tail of eukaryotic mRNA. Ribosomal RNA, transfer RNA, and other non-polyadenylated RNAs are removed by washing under high-salt conditions. Purified mRNA is eluted under low-ionic-strength or heated conditions that disrupt A-T hybridization without degrading the mRNA. |
| Detection Method | Qubit RNA HS or RiboGreen for mRNA quantitation; Bioanalyzer/TapeStation for RNA integrity and size distribution; RT-qPCR of polyadenylated reference genes (GAPDH, ACTB); RNA-seq for transcriptome complexity and coverage analysis; NanoDrop for purity assessment; Agilent mRNA assay for enrichment verification |
| Sample Type | Total RNA extracted from eukaryotic cells and tissues (mammalian, insect, plant, yeast); direct capture from cell lysates using appropriate lysis/binding buffer; total RNA in nuclease-free water or TE buffer; purified total RNA should be of reasonable quality (RIN >5 recommended) |
| Performance Range / Specifications | Bead diameter: 1-2 µm; oligo-dT density: 100-500 pmol dT(25)/mg beads; mRNA binding capacity: 1-3 µg mRNA per mg beads; specificity: >90% mRNA enrichment (assessed by rRNA depletion); rRNA carryover: <10%; elution efficiency: >80% with 10 mM Tris-HCl pH 7.5 at 65-70°C; magnetic separation: <15 seconds |
| Sensitivity / LOD | mRNA capture from as little as 100 ng total RNA input; detection of low-abundance transcripts (<1 copy per cell) after mRNA enrichment; effective poly(A)+ enrichment from partially degraded RNA samples (RIN 5-7) |
| Specificity | Specific hybridization to poly(A) tails (minimum 10-15 A residues required); no binding of rRNA (18S, 28S, 5S, 5.8S), tRNA, or small nuclear RNAs; minimal capture of mRNAs with short (<15 nucleotide) poly(A) tails; >10-fold enrichment of poly(A)+ sequences over total RNA; no capture of prokaryotic mRNA (lacks poly(A)-tail length sufficient for binding) |
| Reaction Conditions / Protocol | For total RNA input: dilute total RNA in binding buffer (20 mM Tris-HCl pH 7.5, 1 M LiCl or 0.5 M NaCl, 2 mM EDTA); heat at 65°C for 2 minutes to denature secondary structure; add pre-equilibrated oligo-dT beads; incubate at room temperature for 5-10 minutes with gentle mixing; magnetically separate; wash 2-3 times with binding buffer; wash once with low-salt buffer; elute mRNA in 10 mM Tris-HCl pH 7.5 at 65-70°C for 2-5 minutes; separate beads; collect mRNA eluate; total protocol: 20-30 minutes; for direct lysis, use supplied lysis/binding buffer |
| Components / Formulation | Oligo-dT(25) functionalized superparamagnetic beads, 10-50 mg/mL in PBS pH 7.4 with 0.02% sodium azide; binding buffer (Tris-HCl, LiCl/NaCl, EDTA) may be included or user-prepared; elution buffer (Tris-HCl) may be included; nuclease-free water not included |
| Storage Conditions | 2-8°C (do not freeze; freezing may denature oligo-dT or cause bead aggregation); protect from light; store in RNase-free conditions; use RNase-free technique when handling; oligo-dT is stable but avoid nuclease contamination |
| Shelf Life | 18 months at 2-8°C; oligo-dT integrity maintained; mRNA binding capacity >85% at 18 months; no RNase contamination throughout shelf life; opened: 6 months with RNase-free handling |
| Package Specifications | 0.5 mL, 1 mL, 5 mL at 10-50 mg/mL; sufficient for 10-100 mRNA isolations; bulk and OEM packaging; trial packs for evaluation |
| Product Form | Dark brown to black suspension in PBS/azide; homogeneous after vortexing; clear supernatant after magnetic separation; oligo-dT validated by hybridization efficiency testing |
| Quality Control | Per lot: oligo-dT density, mRNA binding capacity (HeLa total RNA standard), rRNA depletion efficiency, oligo-dT leaching, mRNA integrity after purification (Bioanalyzer), RNase testing, DNase testing, endotoxin, sterility, particle size |
| Key Features | High-density oligo-dT(25) for efficient poly(A)+ capture; short 20-minute protocol; no columns or centrifugation; mild elution preserves mRNA integrity; compatible with direct lysis for rapid processing; validated for RNA-seq and cDNA synthesis; RNase-free manufacturing |
| Purity | Oligo-dT coupling >90%; endotoxin <0.1 EU/mg; RNase negative; DNase negative; mRNA purity >90% polyadenylated; rRNA in eluate <10% |
| Concentration | 10-50 mg/mL; oligo-dT 100-500 pmol/mg; mRNA binding 1-3 µg/mg; exact values per lot |
| Activity / Unit Definition | mRNA binding: 1-3 µg/mg beads; rRNA reduction >10-fold; elution >80% at 65-70°C; binding linear with total RNA input from 0.1-100 µg |
| Molecular Weight | Oligo-dT(25): ~7,600 Da; each bead carries ~10^5-10^6 oligo-dT molecules; bead molecular weight not defined |
| Source / Origin | Synthetic 5'-amino-modified oligo-dT(25) manufactured by solid-phase phosphoramidite synthesis; HPLC purified; covalently coupled to synthetic polymer-magnetite beads; all components synthetic; RNase-free certified; DEPC-treated process water |
| pH Range / Optimal pH | Hybridization: pH 6.5-8.0 (optimal 7.0-7.5); elution: pH 7.0-8.0; oligo-dT stability: pH 4-10 (short exposure), pH 6-8 (long-term); bead stability: pH 3-12; avoid strong alkaline conditions (pH >12, oligo-dT hydrolysis) |
| Shipping Conditions | Ambient temperature with ice packs for summer; RNase-free packaging; non-hazardous; standard courier; include desiccant for humidity protection |
| Expiration Date / Stability | 18 months at 2-8°C; oligo-dT integrity confirmed by hybridization assay; no nuclease contamination; mRNA binding stable; real-time stability testing at 0, 6, 12, 18 months |
| Regulatory / Compliance | For research use only; not for diagnostic procedures; RNase-free certified; ISO 9001 manufacturing; certificate of analysis per lot |
| Compatibility | Compatible with common lysis/binding buffers: LiCl-based or NaCl-based; works with TRIzol-extracted and column-purified total RNA; mRNA suitable for RNA-seq library preparation (Illumina, PacBio, Nanopore), cDNA synthesis, RT-qPCR, and Northern blotting; compatible with KingFisher and Maxwell automated platforms |
| Recommended Buffer System | Storage: PBS pH 7.4, 0.02% NaN3; binding: 20 mM Tris-HCl pH 7.5, 0.5-1 M LiCl or NaCl, 2 mM EDTA; wash 1: binding buffer; wash 2: 10 mM Tris-HCl pH 7.5, 0.15 M LiCl or NaCl; elution: 10 mM Tris-HCl pH 7.5 (heated to 65-70°C) |
| Application Notes / Precautions | Use RNase-free technique throughout; pre-heat elution buffer to 65-70°C; heat-denature total RNA at 65°C for 2 minutes before binding to disrupt secondary structure; do not exceed 3 µg mRNA loading per mg beads; for input RNA <500 ng, reduce bead volume proportionally; second round of poly(A)+ selection further reduces rRNA contamination; lithium chloride-based buffers provide more stringent A-T hybridization than NaCl-based; avoid repeated freeze-thaw of total RNA input |
| Batch-to-Batch Consistency | Oligo-dT density CV <18%; mRNA binding CV <15%; rRNA depletion consistent across lots; oligo-dT length and purity verified per synthesis batch; RNase testing per lot; hybridization efficiency validated per lot |
For research use only, not for clinical use.
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