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| Product Name | 2′-O-Methyl RNA Oligonucleotide Synthesis Service |
| Catalog No. | PRIPRO-0017 |
| Description | A custom synthesis service for 2′-O-methyl (2′-OMe) modified RNA oligonucleotides. The 2′-OMe modification replaces the 2′-hydroxyl group of ribose with a methoxy group (-OCH₃), creating an RNA analog with dramatically enhanced nuclease resistance and increased RNA binding affinity compared to unmodified RNA. 2′-OMe oligonucleotides form more stable duplexes with complementary RNA targets (Tm increase of 1–3 °C per modified nucleotide) and are resistant to RNase A and other single-strand-specific ribonucleases. The synthesis employs 2′-OMe phosphoramidite building blocks on automated synthesizers, followed by deprotection under mild conditions to preserve the sensitive 2′-OMe functionality. HPLC or PAGE purification is applied based on length and modification density. Each oligo is verified by mass spectrometry. |
| Intended Use | Designed for antisense oligonucleotide studies (splice modulation, translation blocking), siRNA/miRNA inhibitor (antagomir) design, aptamer stabilization, RNA FISH probes, RNase protection assays, and structural biology studies of RNA-protein interactions. |
| Principle / Technology | Solid-phase synthesis using 2′-OMe phosphoramidite monomers; 2′-OMe modification mimics the RNA A-form helix geometry while providing nuclease resistance via steric and electronic effects |
| Detection Method | HPLC or PAGE purity analysis; MALDI-TOF or ESI-MS mass confirmation; thermal denaturation (Tm measurement) to confirm enhanced RNA binding; RNase A resistance assay |
| Sample Type | Complementary RNA targets for hybridization; cellular RNA for antisense targeting; recombinant proteins for binding studies |
| Performance Range / Specifications | 5–50 bases; fully 2′-OMe modified or mixed DNA/2′-OMe chimera; Tm increase 1–3 °C per 2′-OMe residue versus DNA counterpart |
| Sensitivity / LOD | Enhanced detection sensitivity in RNA blot and FISH applications due to increased probe stability; nuclease resistance >48 h in cell culture medium |
| Specificity | 2′-OMe modification enhances Watson-Crick base pairing with RNA; increased binding discrimination against mismatched targets; minimal off-target effects observed |
| Reaction Conditions / Protocol | Dissolve in nuclease-free water or RNase-free TE buffer. For antisense applications, deliver by transfection at 10–100 nM final. For FISH, use 1–10 ng/µL in hybridization buffer. |
| Components / Formulation | Purified 2′-OMe oligonucleotide; COA with MS and purity data; optional endotoxin-free preparation available |
| Storage Conditions | –20 °C or –80 °C; 2′-OMe RNA is significantly more stable than unmodified RNA |
| Shelf Life | 24 months at –20 °C dry; 12 months in solution at –20 °C (RNase-free) |
| Package Specifications | RNase-free screw-cap tube; 10–200 nmol scales; silicone O-ring for seal |
| Product Form | Lyophilized white pellet; RNase-free handling throughout |
| Quality Control | HPLC purity ≥90% (or ≥95% for PAGE); MALDI-TOF MS; DNase and RNase activity testing; endotoxin <0.1 EU/µg optional; Tm measurement with complementary RNA. |
| Key Features | Enhanced RNA binding affinity (1–3 °C Tm increase per residue); RNase A resistant; A-form helix geometry retained; suitable for antisense and splice modulation; MS verified; RNase-free QC; chimera designs available (DNA/2′-OMe mixed). |
| Purity | ≥90% HPLC or ≥95% PAGE; full-length 2′-OMe product |
| Concentration | Resuspend to 100 µM in RNase-free water; A260 quantification with extinction coefficient; scale-dependent yield |
| Activity / Unit Definition | Tm increase 1–3 °C per 2′-OMe residue with complementary RNA; >90% intact after 24 h RNase A treatment; retains ability to block translation or modulate splicing in cellular assays |
| Molecular Weight | Calculated from sequence; 2′-OMe nucleotide approximately 16 Da heavier than corresponding 2′-OH RNA; MS verified |
| Source / Origin | Synthetic 2′-OMe phosphoramidite chemistry; no biological templates |
| pH Range / Optimal pH | Stable pH 5.0–9.0; 2′-OMe linkage stable under standard oligonucleotide handling conditions |
| Shipping Conditions | Ambient temperature as lyophilized pellet; 2′-OMe is far more stable than RNA during transit; cold pack optional |
| Expiration Date / Stability | 24 months at –20 °C dry; 12 months at –20 °C in RNase-free TE or water; stable for multiple freeze-thaw cycles |
| Regulatory / Compliance | For research use only; manufactured in RNase-free facility; suitable as research tool for preclinical oligonucleotide therapeutic studies |
| Compatibility | Compatible with lipid transfection reagents, electroporation, and microinjection; forms stable duplexes with RNA for structural studies; compatible with cellular RNase H when designed as gapmer |
| Recommended Buffer System | RNase-free water or TE buffer (10 mM Tris-HCl pH 7.5, 1 mM EDTA); DEPC-treated water recommended for RNA work |
| Application Notes / Precautions | For antisense splice modulation, design targeting exon-intron junctions or exonic splicing enhancers. Monitor cellular uptake by fluorescence microscopy when using fluorescently labeled 2′-OMe oligos. Verify binding to target by gel-shift assay. |
| Batch-to-Batch Consistency | Mass accuracy ±0.05% by MS; HPLC purity within ±2%; Tm shift within ±1 °C per 2′-OMe residue across batches |
For research use only, not for clinical use.
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