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| Product Name | Locked Nucleic Acid (LNA) Modified Oligonucleotide Synthesis Service |
| Catalog No. | PRIPRO-0018 |
| Description | A custom oligonucleotide synthesis service incorporating locked nucleic acid (LNA) monomers into DNA or RNA oligonucleotides. LNA is a bicyclic RNA analog in which the ribose moiety is chemically locked in the C3′-endo (A-form) conformation by a methylene bridge connecting the 2′-oxygen to the 4′-carbon. This conformational restriction pre-organizes the base for Watson-Crick hybridization, resulting in unprecedented thermal stability increases of 3–8 °C per LNA incorporation with complementary DNA and 5–10 °C per LNA with complementary RNA. LNA oligonucleotides exhibit exceptional single-nucleotide discrimination, making them the gold standard for allele-specific PCR, SNP genotyping probes, and miRNA detection. The synthesis uses LNA phosphoramidite monomers with HPLC purification and rigorous MS quality control. |
| Intended Use | Specifically designed for allele-specific PCR and SNP genotyping with superior mismatch discrimination, microRNA (miRNA) detection by qPCR and Northern blotting, antisense oligonucleotide (gapmer) design with LNA wings, in situ hybridization probes, aptamer affinity maturation, and splice-switching oligonucleotides. |
| Principle / Technology | Incorporation of LNA phosphoramidite monomers during solid-phase synthesis; the methylene bridge locks ribose in C3′-endo, reducing entropy penalty upon hybridization and dramatically increasing thermal stability and mismatch discrimination |
| Detection Method | HPLC purification; MALDI-TOF MS; thermal denaturation analysis (Tm measurement); single-base mismatch discrimination testing; residual phosphoramidite analysis |
| Sample Type | Genomic DNA for SNP genotyping; total RNA or enriched small RNA for miRNA detection; cellular RNA for in situ hybridization |
| Performance Range / Specifications | 8–30 bases standard; LNA content 20–70% of total residues; Tm increase 3–8 °C per LNA versus DNA; optimal for short probes (14–20 nt) targeting AT-rich or difficult sequences |
| Sensitivity / LOD | miRNA detection down to femtomolar range with LNA-enhanced qPCR probes; single-copy SNP detection in genomic DNA with LNA allele-specific primers |
| Specificity | Single-nucleotide discrimination: ΔTm 8–15 °C between perfect match and single mismatch (versus 2–5 °C for DNA); superior to all unmodified oligonucleotide chemistries for allele selectivity |
| Reaction Conditions / Protocol | Design LNA mixmer with LNA spaced every 2–3 DNA residues. Use at 50–200 nM for qPCR probes, 100–500 nM for antisense gapmers. Adjust annealing temperature upward by 2–5 °C per LNA residue. |
| Components / Formulation | HPLC-purified LNA-modified oligo; design consultation report; COA with MS and Tm data; optional endotoxin testing |
| Storage Conditions | –20 °C; LNA is chemically stable; protect from light if dye-labeled |
| Shelf Life | 24 months at –20 °C dry; 12 months in solution at –20 °C |
| Package Specifications | Screw-cap tube; 5–100 nmol scales; custom larger scales by quotation; COA with full characterization |
| Product Form | Lyophilized white pellet |
| Quality Control | HPLC purity ≥90%; MALDI-TOF MS mass confirmation; Tm measurement with complementary DNA and RNA; single-base mismatch discrimination ΔTm reported; residual synthesis reagents below limit. |
| Key Features | Unmatched thermal stability (3–8 °C Tm increase per LNA); superior single-base discrimination (ΔTm 8–15 °C mismatch vs match); ideal for short probes and miRNA detection; enhanced nuclease resistance; compatible with all standard molecular biology workflows; design support included. |
| Purity | ≥90% by HPLC; LNA-containing full-length product |
| Concentration | Resuspend to 100 µM; A260 quantification with LNA extinction coefficient correction; yield reported per scale |
| Activity / Unit Definition | Tm increase 3–8 °C per LNA (vs complementary DNA); 5–10 °C per LNA (vs complementary RNA); mismatch ΔTm 8–15 °C; functional in qPCR, antisense, and hybridization applications |
| Molecular Weight | Calculated from sequence; LNA monomer approximately 16 Da heavier than DNA monomer; verified by MS |
| Source / Origin | Synthetic LNA phosphoramidite chemistry; no biological sources |
| pH Range / Optimal pH | Stable pH 5.0–9.0; LNA linkage chemically robust under all standard oligonucleotide handling conditions |
| Shipping Conditions | Ambient temperature as dry pellet; stable during transit |
| Expiration Date / Stability | 24 months dry at –20 °C; 12 months in solution at –20 °C; LNA is among the most chemically stable nucleic acid modifications |
| Regulatory / Compliance | For research use only; manufactured under ISO 9001 quality system; suitable for preclinical research; GMP production available for therapeutic development upon request |
| Compatibility | Compatible with all DNA and RNA polymerases (adjust extension temperature as needed); compatible with qPCR master mixes; works with standard transfection and hybridization protocols |
| Recommended Buffer System | Nuclease-free water or TE buffer pH 7.5–8.0; standard hybridization buffers with or without formamide |
| Application Notes / Precautions | LNA gapmer design (LNA wings with DNA core) enables RNase H recruitment for antisense applications. For miRNA qPCR, use 3–5 LNA residues in a short (14–18 nt) probe. Avoid self-complementary LNA stretches to prevent aggregation. Order with HPLC purification for best single-base discrimination. |
| Batch-to-Batch Consistency | Tm values consistent within ±1 °C; mismatch ΔTm within ±2 °C; MS mass accuracy ±0.05% across all production lots |
For research use only, not for clinical use.
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