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SP6 RNA Polymerase / 1KU / Cat.#3009
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  SP6 RNA Polymerase

    

#3009

 

500 units

#3009L

 

2,500 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 

¡Ü Preparation of RNA probe for hybridization. 

¡Ü Generation  of  RNA  substrates for studies of RNA processing. 

¡Ü 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¡É. 

 

 

 

STANDARD UNIT ASSAY CONDITIONS 

40mM Tris-HCl (pH7.9), 6mM MgCl2, 10mM DTT, 2mM spemidine, 0.5mM ATP, GTP, CTP, and UTP, 0.5¥ìCi [3H-CTP], and 1¥ìg SP6 promoter-containing plasmid. 

 

CONCENTRATION: 10-20 units/¥ìl. 

 

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¡É. 

 

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. 

 

 

 


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