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 ³²P, ³³P, ³H and ³⁵S 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 MgCl₂, 10mM spermidine, 50mM NaCl. The polymerase is offered as a high-concentration enzyme at 80u/µl (Cat.# P4084).

Choose Your Configuration: Learn more about our custom options for this product at: www.promega.com/custom/

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. Butler, E.T. and Chamberlin, M.J. (1982) J. Biol. Chem. 257, 5772–8.
2. 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 MgCl₂, 10mM DTT, 10mM NaCl, 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.