T7 RNA polymerase  (Synonyms: T7 RNA聚合酶)

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].

产品信息
本品为大肠杆菌重组表达得到。
储存液：20 mM Tris-HCl, 100 mM NaCl, 10 mM DTT, 0.1%Triton X-100, 1 mM EDTA, 50%Glycerol, pH 7.9。
反应液：400 mM Tris-HCl, 60 mM MgCl2, 10 mM DTT, 20 mM spermidine, pH 7.9

使用说明
1. 可参考以下反应体系
0.4 μL 25mM NTP Mix，0.2-1 μg DNA 模板，2 μL 反应缓冲液，0.5 μL RNase Inhibitor (optional, 1 U/μL)，1 μL 0.1 M DTT (optional)，2 μL T7 RNA 聚合酶，使用 RNase Free 水补足体积至 20 μL。
2. 反应条件
37℃ 孵育 0.5-1h，对于小于 300 nt 的片段，建议 37℃ 孵育 2-16 h。

注意事项
1. 反应结束后，使用 2 U DNase Ⅰ（无 RNase）37℃ 15min 去除 DNA 模板。
2. 在 20 μL 反应体系中加入 0.02 U 热稳定无机焦磷酸酶可显著提高转录产量。

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

9014-24-8

液体

Colorless to light yellow

T7 RNA聚合酶

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]