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 purified from a recombinant E.coli strain.
Concentration & Storage Condition- 100 unit/㎕. Store at -20℃. Storage Buffer- 20 mM potassium phosphate, pH 7.7, 1 mM EDTA, 10 mM DTT, 100 mM NaCI, 0.1% (v/v) Triton X-100, 50% (v/v) glycerol. 5x Reaction Buffer- 200 mM Tris-HCl, pH 7.9, 30 mM MgCl₂, 10 mM spermidine, 50 mM NaCl. Unit Definition- One unit is defined as the amount of enzyme required to catalyze the incorporation of 5 nmole of CTP into acid-insoluble product in 60 min at 37°C in a total volume of 100 ㎕.The reaction conditions are: 40 mM Tris-HCl, pH 7.9, 6 mM MgCl₂, 10 mM DTT, 10 mM NaCl, 2 mM spermidine, 0.05% Tween 20, 0.5 mM each of ATP, GTP, CTP, and UTP, 0.5 mCi [³H]CTP and 2 ㎍ supercoiled vector DNA with T7 promoter. QC Tests
- Activity, exo and endonuclease activity test, RNase, DNase, SDS-PAGE purity, performance tests.