T7 RNA Polymerase with buffer F

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.
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C15010HF-25000U set $560.00 Inquiry
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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 3 mM each ATP, UTP, GTP, CTP, and DNA template containing the
T7 RNA Polymerase promoter. Incubate at 37°C.

Storage Buffer
T7 RNA Polymerase is supplied in 100 mM Tris-HCl (pH 7.9), 20 mM KCl, 1 mM DTT, 1 mM EDTA, 0.1% Triton® X-100 and 50% (v/v)

Stored at -20°C. For optimal storage, aliquot the reaction buffer and DTT reagent into smaller quantities and store at
recommended temperature. For most favorable performance, avoid repeated handling and multiple freeze/thaw cycles.​

1. 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.

2. The volume of T7 RNA Polymerase can be titrated between 1-2 μL in the IVT reaction to optimize your assay.
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 0.5-1 μg  -
10X RNA Polymerase Reaction Buffer F 2 μL 1X
ATP (100 mM) 0.6 μL 3 mM
UTP (100 mM) 0.6 μL 3 mM
CTP (100 mM) 0.6 μL 3 mM
GTP (100 mM) 0.6 μL 3 mM
100 mM DTT 2 μL 10 mM
T7 RNA Polymerase (200 U/μL) 1 μL -
RNase inhibitor (optional) 0.5 μL 1 U/μL
Total reaction volume 20 μL -
2. Incubate at 37°C for 30 minutes to 2 hours.
3. Above reaction mixture may be scaled up or down proportionately.