Pseudouridine-5-triphosphate, trisodium salt solution is one of the most commonly used modified nucleoside triphosphates. It is mostly used as the reaction substrate or coenzyme of enzymes, such as in vitro transcription, RNA amplification, siRNA synthesis, etc. The modified mRNA containing pseudouridine has better nuclease stability and translation characteristics, and reduces its immunogenicity. It has a wide range of applications in the field of therapy and diagnosis.This product is produced in accordance with GMP process requirements and provided in liquid form. Feature Validated, product-specific process and analytical methods Product-specific stability Documentation follows applicable GMP guidelines AOF production process and raw materials (TSE & BSE) Nitrosamine statement Regulatory support documents available Large-scale production Nucleotide in the multiple salt form（Na+, Tris etc) always available to meet different downstream application needs Application Modified RNA synthesis and amplification Building block for in vitro transcription Specification CAS No 1175-34-4 (free acid) Formula C9H12N2Na3O15P3 Molecular Weight 550.10 g/moL Purity (HPLC) ≥ 99% Content 100 mM ± 3 mM Structure Component Components No. Name 10650ES20 10650ES70 10650ES80 10650ES10 10650 Pseudo UTP sodium solution GMP-grade (100 mM) 20 μL 100 μL 1 mL 10 mL Shipping and Storage The product is shipped with dry ice and can be stored at -15℃ ~ -25℃ for two years. Figures Standard RNA Synthesis Figure 1. Standard RNA was synthesized in vitro using T7 RNA synthesis kit. The reaction was incubated in PCR instrument at 37℃ for 2h, and then purified by magnetic beads (Cat#12602). The yield result was analyzed by NanoDrop spectrophotometer as shown in Figure 1. Capped RNA Synthesis Figure 2. Synthesis of capped RNA in vitro. The reaction was incubated in PCR instrument at 37℃ for 2h, and then purified by magnetic beads (Cat#12602). The yield result was assayed by NanoDrop spectrophotometer as shown in Figure 2A. The integrity result was analyzed by capillary electrophoresis as shown in Figure 2B. FAQ Q: The extinction coefficient of UTP at 262 nm is 10,000 Lmol-1cm-1, while that of N1-methyl pseudo-UTP at 271 nm is 8,877 Lmol-1cm-1. The difference between the two is significant. When N1-methyl pseudo-UTP replaces UTP for transcription, the A260/A280 value of the resulting mRNA is less than 2.0. The original formula for calculating mRNA concentration (RNA concentration (ng/uL) = 0D260 x N (dilution factor) x 40) is no longer applicable. Could you please tell me what the appropriate coefficient should be? Are there any relevant research data or literature available for sharing? A: Currently, we haven't investigated whether it affects the calculation. The detection method of NanoDrop is relatively rough. If you want a more precise detection, you can use Qubit to measure the concentration of RNA. These two methods are also two of the most common detection methods available on the market. Q: Since we previously used the UTP from another company, and now we want to replace it with the raw materials from Yisheng, are the usage methods different? A: Generally, a complete substitution approach is adopted. The usage amount of pseudouridine or N1-methylpseudouridine is the same as that of uridine in the original system. Documents: Safety Data Sheet 10650_MSDS_HB220701 Manuals 10650_Manual_HB221110
Related
Related