M.B. Enzymes GmbH

T7 RNA polymerase.

Description

T7 RNA polymerase (EC 2.7.7.6) is a DNA-dependent RNA polymerase with a high specificity for T7 promoter sequence (5´TAATACGACTCACTATA3´). The enzyme is a single polypeptide and is isolated from an overproducing recombinant E. coli strain. RNA produced using the T7 RNA polymerase is biologically active as mRNA and can be accurately spliced.

Unit definition

One unit of the enzyme catalyzes the incorporation of 1 nmol of ³H ATP into acid-insoluble product in 60 min at 37°C.

Reaction buffers

B13 (x 5)

Reaction conditions

10 U T7 RNA polymerase, 40 mM Tris-HCl pH 7.5, 6 mM MgCl₂, 2 mM spermidine-HCl, 10 mM DTT, 1-4 mM NTPs. 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. Under optimal conditions, greater than 700 moles of T7 RNA transcript can be synthesized per mol of DNA template.

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. Stahl, S., Zinn, K. (1981) J. Mol. Biol. 148, 485. 2. Davanloo, P. et al. (1984) Proc. Natl. Acad. Sci. USA 81, 2035.

Catalog #

L5.5

Pack size

2.000 U

T7 RNA polymerase.
Description	T7 RNA polymerase (EC 2.7.7.6) is a DNA-dependent RNA polymerase with a high specificity for T7 promoter sequence (5´TAATACGACTCACTATA3´). The enzyme is a single polypeptide and is isolated from an overproducing recombinant E. coli strain. RNA produced using the T7 RNA polymerase is biologically active as mRNA and can be accurately spliced.
Unit definition	One unit of the enzyme catalyzes the incorporation of 1 nmol of ³H ATP into acid-insoluble product in 60 min at 37°C.
Reaction buffers	B13 (x 5)
Reaction conditions	10 U T7 RNA polymerase, 40 mM Tris-HCl pH 7.5, 6 mM MgCl₂, 2 mM spermidine-HCl, 10 mM DTT, 1-4 mM NTPs. 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. Under optimal conditions, greater than 700 moles of T7 RNA transcript can be synthesized per mol of DNA template.
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. Stahl, S., Zinn, K. (1981) J. Mol. Biol. 148, 485. 2. Davanloo, P. et al. (1984) Proc. Natl. Acad. Sci. USA 81, 2035.
Catalog #	L5.5
Pack size	2.000 U