SP6 RNA Polymerase

#3009

 

1,000 units

#3009L

 

5,000 units

DESCRIPTION
SP6 RNA Polymerase is a DNA-dependent RNA polymerase that recognizes and initiates synthesis of RNA on double-stranded DNA templates that carry the SP6 phage promoter. The polymerase is used in vitro to generate large quantities of RNA complementary to one strand of foreign DNA that has been cloned immediately downstream from the promoter in plasmids specifically designed for this purpose.

SOURCE
Recombinant strain of E. coli.

APPLICATIONS
Preparation of RNA probe for hybridization.
Generation of RNA substrates for studies of RNA processing.
Generation of RNA for in vitro translation.

UNIT DEFINITION
One unit is defined as the amount of enzyme required to catalyze the incorporation of 1nmole of nucleotide triphosphate into acid insoluble product in 60 minutes at 37.

STANDARD UNIT ASSAY CONDITIONS
40mM Tris-HCl (pH7.9), 6mM MgCl2, 10mM DTT, 2mM spemidine, 0.5mM ATP, GTP, CTP, and UTP, 0.5Ci [3H-CTP], and 1g SP6 promoter-containing plasmid.

CONCENTRATION: 10-20 units/l.

10X REACTION BUFFER
400mM Tris-HCl (pH7.9), 60mM MgCl2, 20mM spermidine, 100mM DTT.

STORAGE CONDITIONS
20mM Potassium phosphate (pH7.7), 100mM NaCl, 1mM EDTA, 10mM DTT, 0.1% Triton X-100, and 50% glycerol. Store at -20.

REFERENCES
1. Krieg, P. A. and Melton, D. A. (1987) Methods Enzymol. 155, 397.
2. Butler, E. T. and Chamberlain M. J. (1982) J. Biol. Chem. 257, 5772.