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| Product Name | Phosphorothioate-Modified Oligonucleotide Synthesis Service |
| Catalog No. | PRIPRO-0016 |
| Description | A custom oligonucleotide synthesis service producing phosphorothioate (PS)-modified DNA oligos, in which one of the non-bridging oxygen atoms in the phosphodiester backbone is replaced by a sulfur atom. This modification confers substantial resistance to nuclease degradation by serum and cellular nucleases, dramatically extending oligonucleotide half-life in biological fluids and intracellular environments. The phosphorothioate linkage is introduced during solid-phase synthesis via sulfurization of the phosphite triester intermediate. Options include full PS modification (all linkages), partial PS modification (specified positions), or end-capped (2–5 terminal PS linkages at each end). PS oligos retain RNase H-competent hybridization to complementary RNA, making them useful for antisense applications. HPLC purification resolves the diastereomeric mixture inherent to PS modification. |
| Intended Use | Intended for antisense oligonucleotide (ASO) research, gene silencing studies, aptamer stabilization, in vivo oligonucleotide tracking, splice-switching oligonucleotide studies, and any application requiring nuclease-resistant DNA oligonucleotides. |
| Principle / Technology | Sulfur substitution at non-bridging phosphate oxygen via Beaucage reagent or phenylacetyl disulfide (PADS) sulfurization during solid-phase phosphoramidite synthesis |
| Detection Method | HPLC resolution of PS diastereomers; MALDI-TOF MS; ³¹P NMR for PS linkage confirmation; nuclease resistance assay using snake venom phosphodiesterase (SVPD) |
| Sample Type | Cell culture models for antisense studies; in vivo models for pharmacokinetic analysis; in vitro RNase H cleavage assays |
| Performance Range / Specifications | 2–50 bases; full PS, partial PS, or end-capped; PS content 5–100% of linkages; sulfurization efficiency >99% per linkage step |
| Sensitivity / LOD | Nuclease resistance: half-life >24 h in 10% fetal bovine serum at 37 °C versus <1 h for unmodified DNA; RNase H activity retained for gapmer designs |
| Specificity | PS modification does not alter Watson-Crick base pairing specificity; slightly reduced Tm (~0.5–1.5 °C per PS linkage) versus phosphodiester equivalent |
| Reaction Conditions / Protocol | Resuspend in nuclease-free water or PBS. For cell-based studies, deliver via transfection reagent or gymnotic uptake at 1–10 µM. For in vivo studies, prepare in sterile saline. |
| Components / Formulation | HPLC-purified phosphorothioate oligo; COA with sulfurization efficiency data; optional endotoxin testing report |
| Storage Conditions | –20 °C; PS oligos are more stable than unmodified DNA but still benefit from cold storage |
| Shelf Life | 24 months at –20 °C dry; 12 months at –20 °C in solution |
| Package Specifications | Screw-cap tube with 25 nmol–1 µmol scale yield; COA |
| Product Form | Lyophilized white powder (PS modification not visible) |
| Quality Control | HPLC purity ≥90%; MALDI-TOF MS; ³¹P NMR confirming PS linkage presence; iodine oxidation control to confirm >99% sulfurization per step; nuclease resistance time-course; endotoxin <0.1 EU/µg. |
| Key Features | Dramatically enhanced nuclease resistance; customizable PS placement (full, partial, end-capped); maintains base-pairing specificity; RNase H competent for antisense applications; HPLC purified; MS and ³¹P NMR verified. |
| Purity | ≥90% by HPLC; PS diastereomer mixture resolved |
| Concentration | Resuspend to 100 µM stock based on A260; actual yield reported per synthesis scale |
| Activity / Unit Definition | Nuclease resistance: >95% intact after 4 h in 10% FBS; Tm decrease ~0.5–1.5 °C per PS linkage; RNase H cleavage of complementary RNA retained |
| Molecular Weight | Calculated from sequence with 16 Da additional mass per PS linkage (S replaces O); verified by MS |
| Source / Origin | Synthetic DNA with chemical sulfurization step; no biological source |
| pH Range / Optimal pH | Stable pH 6.0–8.5; PS linkage chemically stable under standard storage and assay conditions |
| Shipping Conditions | Ambient temperature; PS oligos more chemically stable than unmodified DNA |
| Expiration Date / Stability | 24 months at –20 °C dry; 12 months at –20 °C in nuclease-free water; PS modification extends solution stability |
| Regulatory / Compliance | For research use only; suitable as tool compounds for preclinical studies; not for human administration without appropriate regulatory approvals |
| Compatibility | Compatible with standard transfection reagents (Lipofectamine, PEI); suitable for gymnotic delivery to many cell types; compatible with cell culture media and biological fluids |
| Recommended Buffer System | Nuclease-free water, PBS, or physiological saline; avoid divalent cation solutions above 10 mM for long-term storage |
| Application Notes / Precautions | For antisense studies, consider gapmer design with PS wings and central DNA gap for RNase H recruitment. Verify nuclease resistance in relevant biological matrix before in vivo studies. PS oligos may exhibit non-specific protein binding at high concentrations. |
| Batch-to-Batch Consistency | Sulfurization efficiency >99% per step across batches; HPLC purity within ±3%; nuclease resistance profile consistent within ±10% |
For research use only, not for clinical use.
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