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T7 RNA Polymerase

T7 RNA Polymerase

DNA-Dependent RNA Polymerase Specific for T7 Promoter Sequences

  • >90% pure as determined by SDS polyacrylamide gel electrophoresis
  • Free of detectable levels of contaminating RNase and DNase activity (<1% release)
  • Incorporates 32P, 33P, 3H and 35S nucleoside triphosphates

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Description

catalog no

Concentration: 10–20u/µl
Size: 1,000u
Catalog number: P2075

Concentration: 10–20u/µl
Size: 5,000u
Catalog number: P2077

Concentration: 80u/µl
Size: 10,000u
Catalog number: P4074

T7 RNA Polymerase is a DNA-dependent RNA polymerase that exhibits extremely high specificity for its cognate promoter sequences. Only T7 DNA or DNA cloned downstream from a T7 promoter can serve as a template for T7 RNA Polymerase-directed RNA synthesis. T7 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. Cat.# P4074 provides T7 RNA Polymerase at a high concentration (80u/µl).

 

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. Milligan, J.F. et al. (1987) Nucl. Acids. Res. 15, 8783–98.
  2. Milligan, J.F. and Uhlenbeck. O.C. (1989) Meth. Enzymol. 180, 51–62.

Storage Buffer: 20mM potassium phosphate (pH 7.7 at 25°C), 1mM EDTA, 10mM DTT, 100mM NaCI, 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 NaCl2, 2mM spermidine, 0.05% Tween® 20, 0.5mM each of ATP, GTP, CTP, and UTP, 0.5μCi [3H]CTP and 2μg supercoiled pGEM®-5Zf(+) Vector DNA.

Protocols

Complete Protocol

T7 RNA Polymerase Protocol

 

Specifications

What’s in the box?
Concentration: 10–20u/µl
Size: 1,000u

Item / Catlog no: P2075 Part # Size Concentration Available Separately
DTT P117B 1 × 250μl 100mM View Product
Transcription Optimized 5X Buffer P118B 1 × 500μl View Product
T7 RNA Polymerase P207B 1 × 1,000u

Use Restrictions

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

Storage Conditions

BB

 

What’s in the box?
Concentration: 10–20u/µl
Size: 5,000u

Item / Catlog no: P2077 Part # Size Concentration Available Separately
DTT P117C 1 × 1.25ml 100mM View Product
Transcription Optimized 5X Buffer P118C 1 × 1.25ml View Product
T7 RNA Polymerase P207E 1 × 5,000u

Use Restrictions

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

Storage Conditions

BB

 

What’s in the box?
Concentration: 80u/µl
Size: 10,000u

Item / Catlog no: P4074 Part # Size Concentration Available Separately
DTT P117B 1 × 250μl 100mM View Product
Transcription Optimized 5X Buffer P118B 1 × 500μl View Product
T7 RNA Polymerase (HC) P407A 1 × 10,000u

Use Restrictions

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

Storage Conditions

BB