R0884

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
• CAS Number: 9014-24-8
• Enzyme Commission number: 2.7.7.6 (BRENDA, IUBMB)
• MDL number: MFCD00132197
• UNSPSC Code: 12352204

Properties

• recombinant: expressed in E. coli
• grade: for molecular biology
• form: buffered aqueous solution
• mol wt: 98.8 kDa
• concentration: 10,000-50,000 U/mL
• UniProt accession no.: P00573
• foreign activity: DNase and RNase, none detected
• storage temp.: −20°C
• Gene Information: bacteriophage T7 ... T7p07(1261050)

Description

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.

Safety Information

• Storage Class: 10 - Combustible liquids
• wgk_germany: WGK 3
• flash_point_f: Not applicable
• flash_point_c: Not applicable