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| Product Name | Phosphorothioate-Modified Antisense Oligonucleotide Synthesis Service |
| Catalog No. | PRIPRO-0026 |
| Description | A custom oligonucleotide synthesis service providing antisense oligos with full phosphorothioate backbone modification for enhanced nuclease resistance. Phosphorothioate internucleotide linkages replace one non-bridging oxygen atom with sulfur at each phosphate, dramatically increasing serum stability while maintaining RNase H-mediated target mRNA cleavage activity. This modification is standard for gapmer antisense oligonucleotides used in gene knockdown studies. |
| Intended Use | Gene expression knockdown through antisense oligonucleotide-mediated RNase H cleavage of target mRNA in cell culture and in vivo models. |
| Principle / Technology | Steric blocking and RNase H recruitment by phosphorothioate-modified DNA oligonucleotides complementary to target mRNA |
| Detection Method | qRT-PCR quantification of target mRNA levels; Western blot for protein knockdown confirmation |
| Sample Type | Transfected cultured cells; systemically or locally administered in animal models |
| Performance Range / Specifications | Synthesis scale: 50 nmol to 10 µmol; length: 15–30 nucleotides; full phosphorothioate backbone; purity: ≥90% by HPLC |
| Sensitivity / LOD | Knockdown of target mRNA ≥70% at 50–100 nM oligonucleotide concentration in standard transfection conditions |
| Specificity | Sequence-dependent; specificity confirmed by mismatch control oligonucleotides |
| Reaction Conditions / Protocol | Design antisense sequence targeting mRNA coding region or 5′-UTR; avoid sequences with strong secondary structure; HPLC purification recommended for in vivo applications |
| Components / Formulation | Lyophilized phosphorothioate oligonucleotide, desalted or HPLC purified per specification; QC certificate with MALDI-TOF mass confirmation and purity assessment |
| Storage Conditions | Lyophilized: –20 °C; reconstituted in nuclease-free water: –20 °C single-use aliquots |
| Shelf Life | 24 months lyophilized at –20 °C |
| Package Specifications | Per oligo: 50 nmol, 200 nmol, 1 µmol synthesis scale |
| Product Form | Lyophilized solid; reconstitute in nuclease-free water or TE buffer |
| Quality Control | MALDI-TOF mass confirmation of full-length product within 0.1% of theoretical mass; HPLC purity as specified |
| Key Features | Full phosphorothioate modification provides extended in vivo half-life from minutes to hours compared to unmodified DNA |
| Purity | HPLC purification ≥90% full-length product; desalted ≥85% |
| Concentration | As specified in synthesis report; typically lyophilized or at 100 µM in TE |
| Activity / Unit Definition | Primer efficiency 90–110% in qPCR; Tm within ±2 °C of calculated value |
| Molecular Weight | As specified per MALDI-TOF certificate; within 0.1% of theoretical mass |
| Source / Origin | Solid-phase phosphoramidite chemical synthesis |
| pH Range / Optimal pH | Stable in TE buffer pH 7.0–8.0; avoid acidic conditions causing depurination |
| Shipping Conditions | Ambient temperature for lyophilized oligos; cold packs for fluorescent probes |
| Expiration Date / Stability | 24 months lyophilized at –20 °C; 6 months reconstituted at –20 °C |
| Regulatory / Compliance | Research use as standard; GMP-grade with full documentation available |
| Compatibility | Compatible with standard PCR, qPCR, and sequencing platforms |
| Recommended Buffer System | Resuspend in nuclease-free water or TE buffer pH 8.0 |
| Application Notes / Precautions | Protect fluorescent probes from light; avoid repeated freeze-thaw cycles; aliquot for single use |
| Batch-to-Batch Consistency | MALDI-TOF mass confirmation per lot; qPCR performance within 0.5 Ct of reference lot |
For research use only, not for clinical use.
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