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T3 RNA Polymerase

DNA-Dependent RNA Polymerase Specific for T3 Promoter Sequences

  • Exhibits extremely high affinity and specificity for T3 promoter sequences
  • >90% pure as determined by SDS polyacrylamide gel electrophoresis
  • Free of detectable levels of contaminating RNase and DNase activity (<1% release)
  • Supplied at a concentration of 10–20u/μl (Cat.# P2083) and 80u/µl (Cat.# P4024)

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Description

catalog no

Size: 1,000u
Catalog number: P2083

Size: 2,500u
Catalog number: P4024

T3 RNA Polymerase is a DNA-dependent RNA polymerase that synthesizes RNA using only T3 DNA or DNA templates cloned downstream from a T3 promoter. The polymerase can incorporate 32P, 33P, 3H and 35S nucleoside triphosphates during transcription. T3 RNA Polymerase is provided with 100mM DTT and Transcription Optimized 5X Buffer: 200mM Tris-HCl (pH 7.9 at 25°C), 30mM MgCl2, 10mM spermidine, 50mM NaCl.

Applications

  • Generation of templates for in vitro translation.
  • Generation of probes for nucleic acid hybridizations.
  • Generation of RNA processing substrates.
  • Generation of antisense RNA.

References

  1. Butler, E.T. and Chamberlin, M.J. (1982) J. Biol. Chem. 257, 5772–8.
  2. Melton, D.A. et al. (1984) Nucl. Acids Res. 12, 7035–56.

Storage Buffer: 20mM potassium phosphate (pH 7.7 at 25°C), 1.0mM EDTA, 10mM DTT, 100mM NaCl, 0.1% (v/v) Triton® X-100 and 50% (v/v) glycerol.

Source: Recombinant E. coli strain.

QC Tests: Activity, SDS-PAGE/purity, DNase, RNase, endonuclease, transcription.

Unit Definition: One unit is defined as the amount of enzyme required to catalyze the incorporation of 5nmol of CTP into acid-insoluble product in 60 minutes at 37°C in a total volume of 100μl. The reaction conditions are: 40mM Tris-HCl (pH 7.9 at 25°C), 6mM MgCl2, 10mM DTT, 10mM NaCl, 2mM spermidine, 0.05% Tween® 20, 0.5mM each of ATP, GTP, CTP, and UTP, 0.5μCi [3H]CTP and 2μg supercoiled pSP6/T3 vector DNA.

Protocols

Complete Protocol

T3 RNA Polymerase Protocol
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What’s in the box?
Size: 1,000u

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Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

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What’s in the box?
Size: 2,500u

[wpdatatable id=364 table_view=regular]

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

BB