Bacteriophage T7 RNA Polymerase is a DNA-dependent RNA polymerase with high specificity for the T7 promoter. This enzyme catalyzes the 5’ to 3’ synthesis of RNA from DNA downstream from its promoter.
Form:
Liquid
Activity:
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. 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:
In 100 mM Tris-HCl, 20 mM KCl, pH 7.9 (1 mM DTT, 1 mM EDTA, 0.1% Triton? X-100 and 50% (v/v) glycerol)
Storage Instruction:
Store at -20°C. Avoid repeated freezing and thawing.
Note:
Transcription reaction should be performed under RNase free condition. Use nucleasefree tubes, reagents, and water to avoid RNase contamination. Also, wear gloves when working with RNA. To obtain optimal condition, NTP concentration can be titrated between 10 – 15 mM. The volume of T7 RNA Polymerase can be titrated between 1-2 uL in the IVT reaction to optimize your assay.