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R0884

Sigma-Aldrich

T7 RNA Polymerase

recombinant, expressed in E. coli, buffered aqueous solution

Synonym(s):

RNA Polymerase T7, RNA Polymerase, T7 from E. coli HMS 174/pAR1219

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About This Item

CAS Number:
EC Number:
MDL number:
UNSPSC Code:
12352204
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recombinant

expressed in E. coli

grade

Molecular Biology

form

buffered aqueous solution

mol wt

98.8 kDa

concentration

10,000-50,000 U/mL

UniProt accession no.

foreign activity

DNase and RNase, none detected

storage temp.

−20°C

Gene Information

bacteriophage T7 ... T7p07(1261050)

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General description

T7 RNA polymerase is highly specific for the bacteriophage T7 promoter and terminator sequences. It is extensively used to prepare RNA transcripts for stuctural and metabolic studies. The RNA transcripts can be converted to probes for sensitive hybridization detection studies. T7 polymerase and dideoxynucleotides can be used to directly sequence DNA.

Components

T7 RNA Polymerase is supplied as a solution of 100 mM NaCl, 50 mM Tris-HCl (pH 7.9), 0.1 mM EDTA, 0.1% Triton X-100, 1 mM DTT, and 50% (v/v) glycerol.

Unit Definition

One unit will catalyze the incorporation of 1 nmol of rNTP into acid-precipitable material in 60 min at 37°C.

Analysis Note

Activity assay: 40 mM Tris-HCl, pH 7.9, 6 mM MgCl2, 4 mM spermidine, 10 mM DTT, 0.5 μM each rNTP + 10 μCi α-32P-UTP, 3-10 units of enzyme, and 1 μg of a 350 bp template are incubated for 10 min at 37°C in a total volume of 100 μl. Typical results are ≥50% incorporation of labeled nucleotide into ≥90% full-length transcript.

Storage Class

10 - Combustible liquids

wgk_germany

WGK 3

flash_point_f

Not applicable

flash_point_c

Not applicable


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Characterization of T7-specific ribonucleic acid polymerase. 1. General properties of the enzymatic reaction and the template specificity of the enzyme.
M Chamberlin et al.
The Journal of biological chemistry, 248(6), 2235-2244 (1973-03-25)
Sangjin Kim et al.
Science (New York, N.Y.), 339(6121), 816-819 (2013-02-16)
Allostery is well documented for proteins but less recognized for DNA-protein interactions. Here, we report that specific binding of a protein on DNA is substantially stabilized or destabilized by another protein bound nearby. The ternary complex's free energy oscillates as
Guillaume Allorent et al.
Plant molecular biology, 82(1-2), 59-70 (2013-03-16)
Arabidopsis seed formation is coupled with two plastid differentiation processes. Chloroplast formation starts during embryogenesis and ends with the maturation phase. It is followed by chloroplast dedifferentiation/degeneration that starts at the end of the maturation phase and leads to the
Wanhui You et al.
Plant molecular biology, 82(1-2), 85-96 (2013-03-21)
RNA-directed DNA methylation (RdDM) is an epigenetic process whereby small interfering RNAs (siRNAs) guide cytosine methylation of homologous DNA sequences. RdDM requires two specialized RNA polymerases: Pol IV transcribes the siRNA precursor whereas Pol V generates scaffold RNAs that interact
William P Robins et al.
Proceedings of the National Academy of Sciences of the United States of America, 110(9), E848-E857 (2013-02-13)
The sequence of a protein determines its function by influencing its folding, structure, and activity. Similarly, the most conserved residues of orthologous and paralogous proteins likely define those most important. The detection of important or essential residues is not always

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