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SP6 RNA Polymerase

SP6 RNA Polymerase

DNA-Dependent RNA Polymerase Specific for SP6 Promoter Sequences

    • Exhibits extremely high affinity and specificity for SP6 promoter sequences
    • >90% pure as determined by SDS polyacrylamide gel electrophoresis
    • Free of detectable levels of contaminating RNase and DNase activity (<1% release)

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Description

catalog no

Concentration: 10–20u/µl
Size: 1,000u
Catalog number: P1085

Concentration: 10–20u/µl
Size: 5,000u
Catalog number: P1081

Concentration: 80u/µl
Size: 2,500u
Catalog number: P4084

SP6 RNA Polymerase is a DNA-dependent RNA polymerase used for in vitro transcription. Only SP6 DNA or DNA cloned downstream from an SP6 promoter can be used as a template for SP6 RNA Polymerase-directed RNA synthesis. The polymerase can incorporate 32P, 33P, 3H and 35S nucleoside triphosphates. SP6 RNA Polymerase is provided with 5X Reaction Buffer: Provided with 100mM DTT and Transcription Optimized 5X Buffer: 200mM Tris-HCl (pH 7.9 at 25°C), 30mM MgCl2, 10mM spermidine, 50mM NaCl. The polymerase is offered as a high-concentration enzyme at 80u/µl (Cat.# P4084).

 

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 buffer (pH 7.7 at 25°C), 1mM 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 pGEM®-5Zf(+) Vector DNA.

Protocols

Complete Protocol

SP6 RNA Polymerase Product Information


Specifications

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

Item / Catlog no: P1085 Part # Size Concentration Available Separately
SP6 RNA Polymerase P108B 1 × 1,000u
DTT P117B 1 × 250μl 100mM View Product
Transcription Optimized 5X Buffer P118B 1 × 500μl View Product

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: P1081 Part # Size Concentration Available Separately
SP6 RNA Polymerase P108G 1 × 5,000u
DTT P117C 1 × 1.25ml 100mM View Product
Transcription Optimized 5X Buffer P118C 2 × 1.25ml View Product

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: P4084 Part # Size Concentration Available Separately
DTT P117B 1 × 250μl 100mM View Product
Transcription Optimized 5X Buffer P118B 1 × 500μl View Product
SP6 RNA Polymerase (HC) P408A 1 × 2,500u

Use Restrictions

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

Storage Conditions

BB