T7 RNA Polymerase (50000 U)

QIAGEN provides a DNA-dependent RNA polymerase with high specificity for the T7 promoter, catalyzing Mg2+-dependent synthesis of RNA from rNTPs in the 5'→3' direction. The enzyme requires its T7 double-stranded DNA promoter to initiate transcription but can transcribe RNA from both single-stranded and double-stranded DNA templates, supporting in vitro transcription and RNA synthesis from a DNA template.

The recombinant enzyme is purified from an E. coli strain expressing the recombinant T7 RNA Polymerase gene, with a molecular weight of 98.9 kDa. One unit is defined as the amount of enzyme that will incorporate 1 nmol of ATP into acid-precipitable material in 1 hour at 37°C. For non-labeled RNA synthesis, a typical reaction uses 100 U of enzyme in a 20 µL volume containing 1X reaction buffer, rNTPs at 500 µM each, and 0.2–1 µg of template DNA, incubated at 37°C for 1 hour.

Quality control testing demonstrates physical purity greater than 99% by SDS-PAGE with silver-stain detection and a specific activity of 312,500 U/mg. Contaminating activities are tested at 500 U: single-stranded exonuclease (<1.0% released), double-stranded exonuclease (<1.0% released), double-stranded endonuclease (no conversion), E. coli DNA contamination (<10 copies), and non-specific RNase (no detectable). The enzyme is supplied in 50 mM Tris-HCl, 100 mM NaCl, 1 mM DTT, 0.1 mM EDTA, 0.1% Triton X-100, 50% glycerol (pH 7.9 at 25°C), and is accompanied by a 10X T7 RNA Polymerase Buffer containing 400 mM Tris-HCl, 60 mM MgCl2, 100 mM DTT, and 20 mM spermidine (pH 7.9 at 25°C). Recommended storage temperature is -25°C to -15°C. The product was developed, manufactured, and sold for in vitro use only.