T7 RNA Polymerase
Source: bacteriophage T7 of Escherichia coli
Modified T7 RNA Polymerase with higher tolerance towards modified nucleotides. Extremely useful for radioactive and non radioactive labeling as well as for RNA synthesis for preparive scale.
Description:
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DNA-dependent RNA polymerase which has stringent specificity for T7 phage promoters sequence (1).
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Ultrapure recombinant enzyme.
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Efficiently synthesizes in vitro transcripts from almost any DNA that is downstream from a T7 promoter (2).
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Suitable for preparing labeled single-stranded RNA probes of high specific activity(3).
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Transcripts can be used as hybridization probes, templates for in vitro translation, substrates in RNA processing systems, or exon and intron mapping of genomic DNA.
Unit Definition: One unit is the amount of enzyme required to incorporate 1 nmol of labeled UTP into acid-soluble material in 1 hr at 37°C.
Storage Conditions: Store at –20°C.
Storage Buffer: 20 mM potassium phosphate (pH 7.5), 0.1 M NaCl, 1 mM EDTA, 1 mM dithiothreitol, 100 µg/ml bovine serum albumin and 50% (v/v) glycerol.
Quality Control: All preparations are assayed for contaminating exonuclease, endonuclease and nonspecific RNase and single- and double-stranded DNase activities. Typical preparations are greater than 90% pure, as judged by SDS polyacrylamide gel electrophoresis.
References:
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Chamberlin, M. and Ring, J. (1973) J. Biol. Chem. 248, 2235-2244.
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Tabor, S and Richardson, C.C. (1985) Proc. Natl. Acad. Sci. U.S.A. 82, 1074-1078.
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Sambrook, J., Fritsch, E. F. and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual, second edition, pp. 10.27-10.37, Cold Spring Harbour Laboratory, Cold Spring Harbour.