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分子别名
T7 RNAPol、T7 RNAP表达宿主
E.coli分子量
100 kDa (Reducing)
纯度
＞95% by SDS-PAGE活性
50 U/μl性状
Liquid缓冲体系
50 mM Tris-HCl, 100 mM NaCl, 1 mM EDTA, 10 mM DTT, 50% (v/v) Glycerol, 0.1% (w/v) Triton® X-100 pH 7.9 @ 25°C
储存条件
Store at -25 ~ -15℃ for 2 years
文献引用
1. Lawyer F C,Stoffel S,Saiki R K ,et al. Isolation, characterization, and expression in Escherichia coli of the DNA polymerase gene from Thermus aquaticus.[J].Journal of Biological Chemistry, 1989, 264.
2. Eom S H,Steitz T A,Wang J M .Structure of Taq polymerase with DNA at the polymerase active site[J].Nature, 1996, 382(6588):278-281.
Bacteriophage T7 RNA Polymerase is a DNA-dependent RNA polymerase that is highly specific for the T7 phage promoters. The 99 KD enzyme catalyzes in vitro RNA synthesis from a cloned DNA sequence under the T7 promoters. RNA produced using the T7 RNA Polymerase is suitable for many applications in research and biotechnology.
Storage Solution: 50 U/ul T7 RNA Polymerase, 50 mM Tris-HCl, 100 mM NaCl, 1 mM EDTA, 10 mM DTT, 50%(v/v) Glycerol, 0.1% (w/v) Triton® X-100 pH 7.9 @ 25°C
10* Reaction Buffer: 400 mM Tris-HCl, 60 mM MgCl2, 10 mM DTT, 20 mM spermidine (pH 7.9 @ 25°C)
Please avoid repeated freeze-thaw cycles.
- One unit is defined as the amount of enzyme that will incorporate 1 nmol ATP into acid-insoluble material in a total reaction volume of 50 μl in 1 hour at 37°C.
生物活性
The experiment was designed. In a 20µl reaction system (40mM Tris-HCl pH7.9, 2mM Spermidine, 6mM MgCl2, 1mM DTT), PCR products containing T7 Promoter were added as template DNA and incubated at 37℃ for 1h. The reaction was terminated after incubation at 70℃ for 10min, and 10U DNase I was added to digest the template DNA. The RNA transcription product obtained was 541nt. Then 5μl reaction product was removed, 1μl 6X Loading Buffer was added, and 1% agar-gel was used for electrophoresis at room temperature, 1X TAE was used as the electrophoresis solution, 160V for 20min. After electrophoresis, the results were photographed. As shown in the figure, this product has excellent transcription effect.
Marker (100~2000 bp)
Lane 1 Negative Control (negative control with no added enzyme only);
Lane 2 50U UA-T7 RNAP;
Lane 3 50U Competitor N-T7 RNAP;
电泳
2μg (R: reducing condition, N: non-reducing condition).
体积排阻色谱(SEC-HPLC)
95.2%
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