T7 RNA Polymerase

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Source

bacteriophage T7 of Escherichia coli

Reagents Supplied

10X T7 RNA Polymerase Reaction Buffer

Description

• DNA-dependent RNA polymerase which has stringent specificity for T7 phage promoters sequence (1)
• Ultrapure recombinant enzyme
  

Applications

• Efficiently synthesizes in vitro transcripts from almost any DNA that is downstream from a T7 promoter (2)
• Suitable for preparing labeled single-stranded RNA probes of high specific activity (3)
• Transcripts can be used as hybridization probes, templates for in vitro translation, substrates in RNA processing systems, or exon and intron mapping of genomic DNA

Unit Definition

One unit is the amount of enzyme required to incorporate 1 nmol of labeled UTP into acid insoluble material for 60 minutes at 37°C 

Concentration

20 - 25 Units/µl

Storage Buffer

20 mM potassium phosphate (pH 7.5)
100 mM NaCl
1mM dithiothreitol
1 mM EDTA
50% (w/v) glycerol

Assay Conditions

40 mM Tris-HCl (pH 7.9)
8 mM MgCl₂
5 mM dithiothreitol
4 mM spermidine (HCl)3
2.5 µg T7 DNA
0.4 mM each of ATP, CTP, GTP and [α-³²P]UTP
Reaction volume is 50 µl

Storage Conditions

Store at -20°C
Product shipped on Dry Ice

References

(1) Chamberlin, M. and Ring, J. (1973) J. Biol. Chem. 248, 2235-2244
(2) Tabor, S. and Richardson, C.C. (1985) Proc. Natl. Acad. Sci. USA 82, 1074-1078
(3) Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989) Mol. Cloning: A Lab. Manual, Second Ed., pp. 10.27-10.37