M.B. Enzymes GmbH

SP6 RNA polymerase.

Description

SP6 RNA polymerase (EC 2.7.7.6) is a DNA-dependent RNA polymerase that recognizes and initiates synthesis of RNA on dsDNA templates that carry the appropriate SP6 promoter (5´ATTTAGGTGACACTATA3´). The enzyme is a single polypeptide and is isolated from an overproducing recombinant E. coli strain.

Unit definition

One unit of the enzyme catalyzes the incorporation of 1 nmol of ³H ATP into acid-insoluble product in 60 min at 40°C.

Reaction buffers

B13 (x5)

Reaction conditions

10 U SP6 RNA polymerase, 2 µg template DNA, 40 mM Tris-HCl pH 7.5, 6 mM MgCl₂, 2 mM spermidine-HCl, 10 mM DTT, 1 mM rNTPs. Additionally, inorganic pyrophosphotase can be added to a final concentration of 4 U/ml to enhance the polymerization reaction. The enzyme will incorporate ³²P, ³⁵S and ³H nucleoside triphosphates

Storage

Store at -20°C.

Applications

production of antisense RNA; RNA labeling; mRNA generation for in vitro translation.

Quality control

activity, SDS-PAGE purity, absence of RNase and DNase activities, specific performance tests.

References

1. Butler, E.T., Chamberlin, M.J. (1982) J. Biol. Chem. 257, 5772. 2. Melton, D.A. et al. (1984) Nucl. Acids Res. 12, 7035.

Catalog #

L4.5

Pack size

2.000 U

SP6 RNA polymerase.
Description	SP6 RNA polymerase (EC 2.7.7.6) is a DNA-dependent RNA polymerase that recognizes and initiates synthesis of RNA on dsDNA templates that carry the appropriate SP6 promoter (5´ATTTAGGTGACACTATA3´). The enzyme is a single polypeptide and is isolated from an overproducing recombinant E. coli strain.
Unit definition	One unit of the enzyme catalyzes the incorporation of 1 nmol of ³H ATP into acid-insoluble product in 60 min at 40°C.
Reaction buffers	B13 (x5)
Reaction conditions	10 U SP6 RNA polymerase, 2 µg template DNA, 40 mM Tris-HCl pH 7.5, 6 mM MgCl₂, 2 mM spermidine-HCl, 10 mM DTT, 1 mM rNTPs. Additionally, inorganic pyrophosphotase can be added to a final concentration of 4 U/ml to enhance the polymerization reaction. The enzyme will incorporate ³²P, ³⁵S and ³H nucleoside triphosphates
Storage	Store at -20°C.
Applications	production of antisense RNA; RNA labeling; mRNA generation for in vitro translation.
Quality control	activity, SDS-PAGE purity, absence of RNase and DNase activities, specific performance tests.
References	1. Butler, E.T., Chamberlin, M.J. (1982) J. Biol. Chem. 257, 5772. 2. Melton, D.A. et al. (1984) Nucl. Acids Res. 12, 7035.
Catalog #	L4.5
Pack size	2.000 U