This product is a bacteriophage T7 RNA polymerase derived from recombinant expression in E. coli. It is a T7 RNA polymerase obtained by mutation after modification and optimization of wild-type T7, which can significantly reduce the dsRNA content. Double-stranded DNA containing the T7 promoter sequence is used as a template and NTP is used as a substrate to synthesize RNA complementary to the reverse single-stranded DNA downstream of the promoter. Double-stranded linear blunt-end or 5' protruding end DNA can be used as a substrate template for T7 RNA polymerase, so linear plasmids and PCR products can be used as templates for in vitro RNA synthesis. Specifications Source Recombinant E. coli Optimum temperature 37℃ Activity 1000 U/μL Residual RNase Negative Storage buffer 50 mM Tris-HCl, 1 mM EDTA, 10 mM DTT, 100 mM NaCl, 0.1% Triton X-100, 50% (v/v) glycerol, pH7.9@25℃ Activity unit definition The amount of enzyme required to incorporate 1 nmol of [3H] GMP into the acid-insoluble precipitate within 1 hour at 37℃ and pH8.0 is defined as one activity unit. Components Components No. Name 10636ES50 (50 KU) 10636ES60 (500 KU) 10636ES72 (1 MU) 10636ES86 (10 MU) 10636 HieffTM T7 RNA Polymerase (low dsRNA, 1000 U/μL) 50 μL 500 μL 1 mL 10 mL Storage This product should be stored at -25~-15℃ for 1 year. Figures 1. Customer feedback 1 Experimental Design: Parameter Condition Objective To compare application performance between WT T7 and Low-dsRNA T7 Reaction Volume 20 µL Template Length 2 kb Test Samples WT T7 & Low-dsRNA T7 RNA polymerase Reaction Temperature 27 °C Reaction Time 2 h Results: Table 1. Comparison of dsRNA Content and Yield Between WT T7 and Low-dsRNA T7 Enzyme Type dsRNA Content (%) Integrity (%) Reaction Scale WT T7 0.0030% 92.5 180 µg (20 µL) Low-dsRNA T7 0.0003% 93.0 180 µg (20 µL) Result Summary: Low-dsRNA T7 reduced dsRNA content by approximately 10-fold while maintaining comparable RNA yield and integrity relative to WT T7. 2. Customer feedback 2 Table 2. Performance Comparison Under Different GAG Final Concentrations Yield (mg/mL) GAG Final Concentration (mM) Yield (mg/mL) Integrity (%) Capping Efficiency (%) Transfection Efficiency (%) WT Yeasen supplier A* WT Yeasen supplier A* WT Yeasen supplier A* WT Yeasen supplier A* 10 12.14 11.376 12.103 88.1 92.5 92.3 96.7 100 100 44.60 33.63 30.76 5 11.82 10.716 11.337 82.2 91.1 91.5 98.3 100 100 42.30 32.78 32.37 2.5 11.31 10.716 11.066 82.9 91.7 91.8 98.8 97.3 97.8 39.78 31.68 32.59 1.5 11.98 — — 78.9 — — 97.7 — — — — — 1 11.75 — — 80.0 — — 90.5 — — — — — 0.5 11.65 — — 80.9 — — 70.1 — — — — — Result Summary: Effects of cap analog concentration on IVT performance. Reactions were carried out in a 20 µL system at 27 °C for 2 h using different T7 enzymes. RNA yield remained stable as cap analog input decreased, while integrity and capping efficiency showed a gradual decline. A cap analog concentration of ≥2.5 mM is recommended for optimal performance. Documents: Safety Data Sheet 10636_MSDS_HB260115 Manuals 10636_Manual_Ver.EN20251208
