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| Product Name | NTP Mix (25 mM Each, Transcription Grade) |
| Catalog No. | IEC-HMM-0061 |
| Description | An equimolar mixture of ultrapure ribonucleoside triphosphates (ATP, CTP, GTP, UTP) at 25 mM each in nuclease-free water. These nucleotides serve as substrates for bacteriophage RNA polymerases (T7, SP6, T3) in in vitro transcription for synthesis of mRNA, siRNA, guide RNA, and other RNA species. |
| Intended Use | Substrate for RNA polymerases in in vitro transcription reactions for mRNA vaccine production, CRISPR guide RNA synthesis, RNA probe generation, RNA structure studies, and general molecular biology RNA synthesis applications. |
| Principle / Technology | Ribonucleoside triphosphates (NTPs) are incorporated by DNA-dependent RNA polymerases into nascent RNA transcripts through phosphodiester bond formation at the 3'-OH terminus, releasing pyrophosphate. Magnesium ions serve as essential cofactors in the catalytic mechanism. |
| Detection Method | Purity verified by HPLC. Concentration determined by UV spectrophotometry. Functional validation by T7 RNA polymerase in vitro transcription yield assay. |
| Sample Type | Substrate component for in vitro transcription reactions using T7, SP6, or T3 RNA polymerase and double-stranded DNA templates containing the cognate promoter. |
| Performance Range / Specifications | Each NTP: 25 mM +/- 5%. Total nucleotide: 100 mM. Working concentration in transcription: typically 5-7.5 mM each NTP. Supports RNA yields of 100-200 ug per ug template under optimized conditions. |
| Sensitivity / LOD | Supports RNA synthesis from as little as 100 ng template DNA in 20 uL transcription reaction. |
| Specificity | Ultrapure NTPs with no detectable contamination by dNTPs, NDPs, NMPs, or nucleosides that could inhibit transcription or cause premature termination. |
| Reaction Conditions / Protocol | 1x transcription buffer, 5 mM each NTP, 10-20 U T7 polymerase, 0.5-1 ug template DNA, 37 °C 1-4 h. For cap analog co-transcriptional capping, adjust GTP accordingly. |
| Components / Formulation | 25 mM ATP, 25 mM CTP, 25 mM GTP, 25 mM UTP in ultrapure nuclease-free water, pH adjusted to 7.0 with NaOH. |
| Storage Conditions | Store at -20 °C to -80 °C. Aliquot to avoid repeated freeze-thaw and minimize NTP hydrolysis. |
| Shelf Life | 24 months at -20 °C; 36 months at -80 °C. |
| Package Specifications | 0.25 mL, 1 mL, 5 mL, 25 mL. Bulk packaging for manufacturing available on request. |
| Product Form | Clear, colorless aqueous solution. pH neutral. |
| Quality Control | Each lot tested by: HPLC purity (>= 99% per NTP); UV concentration verification; DNase and RNase contamination assays; in vitro transcription functional test (yield >= 100 ug RNA per ug template); transcript integrity by capillary electrophoresis or denaturing gel electrophoresis. |
| Key Features | Supports high-yield in vitro transcription; ultrapure HPLC-verified; nuclease-free certification; compatible with cap analog co-transcriptional capping; suitable for GMP-grade mRNA production with appropriate quality grade selection. |
| Purity | >= 99% per individual NTP by HPLC analysis. |
| Concentration | 25 mM each NTP (100 mM total). Alternative concentrations available (10, 50, 100 mM). |
| Activity / Unit Definition | Rate of incorporation by T7 RNA polymerase: ~200-260 nt/s at 37 °C under saturating NTP and Mg²⁺ conditions. |
| Molecular Weight | ATP: 551.1 g/mol; CTP: 527.1 g/mol; GTP: 567.1 g/mol; UTP: 550.1 g/mol (all as trisodium or disodium salts). Precise molecular weight depends on counterion and hydration state. |
| Source / Origin | Synthesized from nucleoside precursors via phosphorylation. Manufactured in dedicated nucleotide synthesis facilities under stringent quality control. |
| pH Range / Optimal pH | Solution pH 7.0-7.5. Optimal transcription buffer pH: 7.7-8.3 (Tris-based). NTPs are acidic; buffer capacity must be sufficient to maintain pH during reaction. |
| Shipping Conditions | Shipped with cold packs to maintain below 10 °C. For GMP-grade material, ship on dry ice. Stable at ambient temperature for up to 3 days. |
| Expiration Date / Stability | 24 months at -20 °C. Hydrolysis rate increases at alkaline pH and elevated temperatures. Aliquot and store at -80 °C for maximum stability of bulk material. |
| Regulatory / Compliance | ISO 9001 and ISO 13485 certified. GMP-grade NTPs with full traceability and regulatory documentation available for therapeutic mRNA manufacturing. |
| Compatibility | Compatible with T7, SP6, and T3 RNA polymerases. Not a substrate for DNA polymerases. Compatible with cap analogs (ARCA, CleanCap, m7GpppG) for co-transcriptional capping. Compatible with modified NTPs for functionalized RNA synthesis. |
| Recommended Buffer System | Supplied in nuclease-free water without buffer. Transcription buffer provided separately and must contain Mg²⁺ (typically 15-25 mM final) as essential cofactor. |
| Application Notes / Precautions | Thaw on ice and vortex briefly before use. For large-scale transcription, include inorganic pyrophosphatase to prevent magnesium pyrophosphate precipitation. If NTPs precipitate during storage, warm to room temperature and vortex; precipitation does not affect quality. For GMP applications, use dedicated, qualified aliquots. |
| Batch-to-Batch Consistency | HPLC purity and transcription yield verified per batch. Yield consistent within +/- 15% of reference across production lots. |
For research use only, not for clinical use.
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