T7 RNA polymerase 

T7 RNA polymerase is a polymerase expressed by Escherichia coli from the RNA polymerase gene of T7 bacteriophage. T7 RNA polymerase is highly specific and involved in in vitro transcription (IVT) of mRNA. In the presence of Mg²⁺, T7 RNA polymerase only uses the single-stranded or double-stranded DNA containing the T7 promoter sequence as a template, and uses NTP as a substrate to synthesize RNA complementary to the single-stranded DNA downstream of the promoter^[1][2].

Product information
This product is recombinantly expressed in Escherichia coli .
Storage buffer: 20 mM Tris-HCl, 100 mM NaCl, 10 mM DTT, 0.1% Triton X-100, 1 mM EDTA, 50% Glycerol, pH7.9.
Reaction buffer: 400 mM Tris-HCl, 60 mM MgCl2, 10 mM DTT, 20 mM spermidine, pH 7.9

Instructions
1. Please refer to the following reaction system
0.4 μL 25mM NTP Mix, 0.2-1 ug DNA template, 2 μL reaction buffer, 0.5 μL RNase Inhibitor (optional, 1 U/μL), 1 μL 0.1 M DTT (optional), 2 μL T7 RNA polymerase, use RNase Make up the volume to 20 μL with free water.
2.Reaction conditions
Incubate at 37°C for 0.5-1h. For fragments smaller than 300 nt, it is recommended to incubate at 37°C for 2-16 h.

Notes
1. After the reaction, use 2 U DNase I (RNase-free) at 37°C for 15 minutes to remove the DNA template.
2. Adding 0.02 U thermostable inorganic pyrophosphatase to the 20 μL reaction system can significantly increase the transcription yield.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

9014-24-8

Liquid

Colorless to light yellow

[T7 RNA polymerase]

≥50 U/μL

One unit is defined as the amount of enzyme that catalyze the incorporation of 1 nmol AMP into the polynucleotide at 37°C in 60 min.

Room temperature in continental US; may vary elsewhere.

Please store the product under the recommended conditions in the Certificate of Analysis.

• [1]. Dousis A, et al. An engineered T7 RNA polymerase that produces mRNA free of immunostimulatory byproducts. Nat Biotechnol. 2023 Apr;41(4):560-568.  [Content Brief]

  [2]. Borkotoky S, et al. The highly efficient T7 RNA polymerase: A wonder macromolecule in biological realm. Int J Biol Macromol. 2018 Oct 15;118(Pt A):49-56.  [Content Brief]