Promega, P4074, 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 MgCl 2 , 10mM spermidine, 50mM NaCl. Cat.# P4074 provides T7 RNA Polymerase at a high concentration (80u/µl).
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
Milligan, J.F. et al. (1987) Nucl. Acids. Res. 15 , 8783–98.
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 MgCl 2 ,10mM DTT, 10mM NaCl 2 , 2mM spermidine, 0.05% Tween® 20, 0.5mM each of ATP, GTP, CTP, and UTP, 0.5μCi [ 3 H]CTP and 2μg supercoiled pGEM®-5Zf(+) Vector DNA.