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Enzymes for Molecular Biology

Enzymes for Molecular Biology: Enzymes for Molecular Biology

T7 and SP6 RNA Polymerases

T7 and SP6 RNA Polymerases produce defined RNA by in vitro transcription of DNA cloned into a plasmid or other vector downstream from their respective promoter.

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Description

T7 and SP6 T3 RNA Polymerases produce defined RNA by in vitro transcription of DNA cloned into a plasmid or other vector downstream from their respective promoter.

Notes

  1. A buffer package containing 5X Transcription Buffer and DTT may be purchased separately.
  2. For synthesis of large amounts of RNA, use the AmpliScribe™ High Yield Transcription Kits or for synthesis of large amounts of 5'-capped RNA use the AmpliCap™ High Yield Message Maker Kits.
  3. For synthesis of high yields of RNA that is completely resistant to RNase A digestion, use the DuraScribe® T7 Transcription Kit.

Unit Definition

One unit catalyses the incorporation of 1 nmole of a labelled ribonucleoside triphosphate into RNA in 1 hour at 37°C in 40mM Tris-HCl, pH 7.5, 6mM MgCl2, 10mM NaCl, 2mM spermidine, 10mM DTT, and 0.5mM each NTP using a DNA template with the appropriate T7, SP6, or T3 promoter. 

Storage Buffer

50% glycerol containing 50mM Tris-HCl, pH 7.5, 0.1M NaCl, 1.0mM DTT, 0.1mM EDTA, and 0.1% Triton X-100. 

5X Transcription Buffer

0.2M Tris-HCl, pH 7.5, 50 mM NaCl, 30mM MgCl2, and 10mM spermidine. 

Quality Control

 These RNA Polymerases are tested in transcription and are free of detectable exo- and endonuclease and RNase activities, and E. coli RNA polymerase.


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Protocols

Protocols for: T7, T3 & SP6 Phage RNA Polymerases

 http://www.epibio.com/pdftechlit/techlit_t7-t3-sp6_rna_polymerases.asp

T7, T3 & SP6 Phage RNA Polymerases Protocol

(catalogue number for all T7, T3 or SP6 Phage RNA Polymerases) 

Please note: all protocols off site are the responsibility of the products supplier

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References

References

T7 RNA Polymerase

  1. Davanloo, P. et al. (1984) Proc. Natl. Acad. Sci. USA 81, 2035.
  2. Eberwine, J. (1992) Proc. Natl. Acad. Sci. USA 89, 3010.(Note: Use of T7 RNA Polymerase for this application may require a license.)
  3. Stahl, S. J. and Zinn, K. (1981) J. Mol. Biol. 148, 481.

T3 RNA Polymerase

  1. Konarska, M.M. et al. (1984) Cell 38, 731.
  2. Parvin, J.D. et al. (1986) DNA 5, 167.

SP6 RNA Polymerase

  1. Butler, E.T. and Chamberlin, M.J. (1982) J. Biol. Chem. 257

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Notes

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Applications & Benefits

Applications

  • Synthesis of RNA for nucleic acid amplification methods or gene expression studies.
  • RNA synthesized can be used as: a hybridization probe, antisense RNA, a ribozyme, a template for in vitro translation, a precursor mRNA for splicing or other processing studies, or to make dsRNA for RNA interference or gene silencing

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Related products

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