T7 RNA Polymerase

One unit is defined as the amount of enzyme incorporating 1.5 nmol NTP into acid-precipitable material in 60 min at 37°C. The enzyme is assayed in 40 mM Tris-HCl pH 8.0 (at 25°C), 2 mM Spermidine, 10 mM DTT, 6 mM MgCl₂, 2 mM NTPs (0.5 mM each), 1.25 µCi [3H]-GTP (0.25 nmol ~1% of GTPs), and 1 µg SspI-digested pET Blue-1 dsDNA template.

99.97 kDa

EDTA can be used to stop reaction.

The enzyme is stable at -20°C for up to 1 year in the supplied storage buffer. It is recommended to avoid repeated freeze-thaw of formulations not containing glycerol.

100 U T7 RNA Polymerase was incubated with a supercoiled plasmid (1 µg) for 4 hours at 37°C. Agarose gel electrophoresis did not reveal any transformation of closed circular DNA to nicked DNA.

200 U T7 RNA Polymerase was incubated with M13 ssDNA (0.5 µg) for 4 hours at 37°C. Agarose gel electrophoresis did not reveal any visible signs of ssDNA degradation.

200 U T7 RNA Polymerase was incubated with 3H-dATP labelled ds or ssDNA (0.5 µg, 500 bp) for 4 hours at 37°C. Acid soluble radioactivity from labelled DNA was not significant over blank test for either substrate.

200 U T7 RNA Polymerase was incubated with a 2 kb RNA transcript (1 µg) for 4 hours at 37°C. Agarose gel electrophoresis did not reveal any visible signs of RNA degradation.

Protein purity of T7 RNA Polymerase was ≥95% as determined by SDS-PAGE analysis.