T7 RNA Polymerase

T7 RNA Polymerase is a DNA-dependent RNA polymerase that exhibits extremely high specificity for its cognate promoter sequences. Only T7 DNA or DNA cloned downstream from a T7 promoter can serve as a template for T7 RNA Polymerase-directed RNA synthesis. T7 RNA Polymerase is provided with 100mM DTT and Transcription Optimized 5X Buffer: 200mM Tris-HCl (pH 7.9 at 25°C), 30mM MgCl₂, 10mM spermidine, 50mM NaCl. Cat.# P4074 provides T7 RNA Polymerase at a high concentration (80u/µl).

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Applications

• Generation of templates for in vitro translation.
• Generation of probes for nucleic acid hybridizations.
• Generation of RNA processing substrates.
• Generation of antisense RNA.

References

1. Milligan, J.F. et al. (1987) Nucl. Acids. Res. 15, 8783–98.
2. Milligan, J.F. and Uhlenbeck. O.C. (1989) Meth. Enzymol. 180, 51–62.

Storage Buffer: 20mM potassium phosphate (pH 7.7 at 25°C), 1mM EDTA, 10mM DTT, 100mM NaCI, 0.1% (v/v) Triton® X-100 and 50% (v/v) glycerol.

Source: Recombinant E. coli strain.

QC Tests: Activity, SDS-PAGE/purity, DNase, RNase, endonuclease, transcription.

Unit Definition: One unit is defined as the amount of enzyme required to catalyze the incorporation of 5nmol of CTP into acid-insoluble product in 60 minutes at 37°C in a total volume of 100μl. The reaction conditions are: 40mM Tris-HCl (pH 7.9 at 25°C), 6mM MgCl₂,10mM DTT, 10mM NaCl₂, 2mM spermidine, 0.05% Tween® 20, 0.5mM each of ATP, GTP, CTP, and UTP, 0.5μCi [³H]CTP and 2μg supercoiled pGEM®-5Zf(+) Vector DNA.