T3 RNA Polymerase
T3 RNA Polymerase is a DNA-dependent RNA polymerase that synthesizes RNA using only T3 DNA or DNA templates cloned downstream from a T3 promoter. The polymerase can incorporate 32P, 33P, 3H and 35S nucleoside triphosphates during transcription. T3 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.
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- Generation of templates for in vitro translation.
- Generation of probes for nucleic acid hybridizations.
- Generation of RNA processing substrates.
- Generation of antisense RNA.
- Butler, E.T. and Chamberlin, M.J. (1982) J. Biol. Chem. 257, 5772–8.
- Melton, D.A. et al. (1984) Nucl. Acids Res. 12, 7035–56.
Storage Buffer: 20mM potassium phosphate (pH 7.7 at 25°C), 1.0mM EDTA, 10mM DTT, 100mM NaCl, 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 100ul. The reaction conditions are: 40mM Tris-HCl (pH 7.9 at 25°C), 6mM MgCl2, 10mM DTT, 10mM NaCl, 2mM spermidine, 0.05% Tween® 20, 0.5mM each of ATP, GTP, CTP, and UTP, 0.5uCi [3H]CTP and 2ug supercoiled pSP6/T3 vector DNA.
T3 RNA Polymerase Protocol
PDF (113 KB)
Certificate of AnalysisLookup Certificate of Analysis
-30C TO -10C
For Research Use Only. Not for Use in Diagnostic Procedures.