Promega, P4084, SP6 RNA Polymerase

SP6 RNA Polymerase is a DNA-dependent RNA polymerase used for in vitro transcription. Only SP6 DNA or DNA cloned downstream from an SP6 promoter can be used as a template for SP6 RNA Polymerase-directed RNA synthesis. The polymerase can incorporate 32 P, 33 P, 3 H and 35 S nucleoside triphosphates. SP6 RNA Polymerase is provided with 5X Reaction Buffer: 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. The polymerase is offered as a high-concentration enzyme at 80u/µl (Cat.# P4084).
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
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 buffer (pH 7.7 at 25°C), 1mM 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 100μl. The reaction conditions are: 40mM Tris-HCl (pH 7.9 at 25°C), 6mM MgCl 2 , 10mM DTT, 10mM NaCl, 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.