T7 RNA Polymerase

Bacteriophage T7 RNA Polymerase is a DNA-dependent RNA polymerase with high specificity for the T7 promoter. This enzyme catalyzes the 5’→3’synthesis of RNA from DNA downstream from its promoter.
No. Size Price Qty Status
C15010-5000U 5,000 U (50 U/μL) $136.00
C15010-25000U 25,000 U (50 U/μL) $560.00
C15010-50000U 50,000 U (50 U/μL) $1,000.00
C15010H-25000U 25,000 U (200 U/μL) $560.00
C15010-B 1 mL $0.00
The price does not include shipping fee and tax. ORDER
Escherichia coli
>98% as determined by SDS-PAGE. Purified by Ni-NTA chromatography.
Unit Definition:
One unit is defined as the amount of the enzyme incorporates 1 nmol of ATP into acid-insoluble product in 1 hour at 37°C.
Reaction Condition:
1X RNA Polymerase Reaction Buffer, supplemented with 0.5 mM each ATP, UTP, GTP, CTP, and DNA template containing the T7 RNA Polymerase promoter. Incubate at 37°C.
10X RNA Polymerase Reaction Buffer: 400 mM Tris-HCl (pH 8.0), 60 mM MgCl2, 100 mM DTT, and 20 mM spermidine.
Storage Buffer:
T7 RNA Polymerase is supplied in 20 mM potassium phosphate (pH 7.7), 100 mM NaCl, 10 mM DTT, 1 mM EDTA, 0.1% Triton® X-100 and 50% (v/v) glycerol.

Stored at -20°C. Avoid repeated freeze/thaw cycles.

Transcription reaction should be performed under RNase free condition. Use nuclease-free tubes, reagents, and water to avoid RNase contamination. Also, wear gloves when working with RNA.
Standard RNA synthesis procedures:
1. Below reaction mixture should be prepared under room temperature and combined in the following order:
Component Amount Final concentration
Nuclease-Free H2O X μL -
Template DNA 0.2-1 μg 10-50 ng/μL linearized DNA
10X RNA Polymerase Reaction Buffer 2 μL 1X
10 mM ATP 1 μL 0.5 mM
10 mM UTP 1 μL 0.5 mM
10 mM CTP 1 μL 0.5 mM
10 mM GTP 1 μL 0.5 mM
T7 RNA Polymerase (50 U/μL) 1-2 μL -
RNase inhibitor (optional) X μL 20 U
Total reaction volume 20 μL -
2. Incubate at 37°C for 2-16 hours.
3. Above reaction mixture may be scaled up or down proportionately.