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Description
T7 RNA PolymeraseProduct Specification Synonyms T7 RNAPolT7 RNAP Expression System E. coli Molecular Weight 100 kDa (Reducing) Purity 95% by SDS PAGE Physical Appearance Liquid Storage Buffer 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 @ 25C Stability & Storage Store at 25 ~ 15 for 2 years Reference 1. Lawyer F C,Stoffel S,Saiki R K ,et al. Isolation, characterization, and expression in Escherichia coli of
Product Specification
| Synonyms | T7 RNAPol、T7 RNAP |
| Expression System | E.coli |
| Molecular Weight | 100 kDa (Reducing) |
| Purity | ＞95% by SDS-PAGE |
| Physical Appearance | Liquid |
| Storage Buffer | 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 |
| Stability & Storage | Store at -25 ~ -15℃ for 2 years |
| Reference | 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. |
Background
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.
Components
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)
Protocol
1. Assemble the reaction at room temperature in the following order.
Components |
Volume |
Final Concentration |
10X Reaction Buffer |
2 µl |
1 X |
NTP mix, 10 mM each |
1 µl |
0.5 mM each |
RNase Inhibitor（40 U/ul） |
0.5 µl |
1 U/µl |
50 mM DTT |
2 µl |
5 mM |
Template DNA |
variable |
20 ng–1 µg |
T7 RNA Polymerase |
1 µl |
2.5 U/µl |
DEPC-treated Water |
Up to 20µl |
- |
2. Incubate at 37°C for 1 hour. For shorter (< 300 nt) transcripts incubate at 37°C for 2–16 hours.
Guidelines
Please avoid repeated freeze-thaw cycles.
Unit Definition
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