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R0759

Sigma-Aldrich

RNA Polymerase from Salmonella enterica serotype typhimurium, SP6-infected

buffered aqueous solution

Synonym(s):

SP6 RNA Polymerase

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About This Item

CAS Number:
MDL number:
UNSPSC Code:
12352204

grade

for molecular biology

form

buffered aqueous solution

concentration

10000-50000 units/mL

UniProt accession no.

foreign activity

DNase, RNase, protease, none detected
nonspecific endonuclease, absence of activity (after 20 hour incubation)

storage temp.

−20°C

Gene Information

bacteriophage SP6 ... SP6p08(1481778)

General description

Sp6 Polymerase is a DNA-dependent RNA polymerase with high specific activity for the bacteriophage Sp6 promoter. The enzyme is efficient in transcribing large quantities of RNA from the DNA sequence (polylinker) downstream from the promoter. Simultaneous transcription by Sp6 and Sp7 polymerases may occur without interfering with each other.

Application

Suitable for:
  • Synthesis of labeled and unlabeled RNA transcripts
  • RNase protection assays
  • Generating RNA transcripts for in vitro translation
  • Use in in vitro transcription of RNA transcripts, antisense RNA (from reversed cloned DNA insert), and biologically active mRNA

Components

Protease is supplied in a solution of 100 mM NaCl, 50 mM Tris-HCl (pH 7.9), 0.1 mM EDTA, 0.1% Triton X-100, 1 mM DTT, and 50% (v/v) glycerol.

Unit Definition

One unit will catalyze the incorporation of 1 nmol of rNTP into acid-precipitable material in 60 min at 37 °C.

Analysis Note

Activity assay: 40 mM Tris-HCl, pH 7.9, 6 mM MgCl2, 4 mM spermidine, 10 mM DTT, 0.5 μM each rNTP + 10 μCi α-32P-UTP, 3-10 units of enzyme, and 1 μg of a 350 bp template are incubated for 10 min at 37 °C in a total volume of 100 μl. Typical results are ≥50% incorporation of labeled nucleotide into ≥90% full-length transcript.

Other Notes

Sp6 polymerase exhibits 30% more activity at 40°C than at 37°C.

pictograms

Exclamation mark

signalword

Warning

hcodes

Hazard Classifications

Aquatic Chronic 3 - Eye Irrit. 2

Storage Class

10 - Combustible liquids

wgk_germany

WGK 2

flash_point_f

Not applicable

flash_point_c

Not applicable


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Sambrook, J., et al.
Molecular Cloning: A Laboratory Manual, 18-18 (1989)
D A Melton et al.
Nucleic acids research, 12(18), 7035-7056 (1984-09-25)
A simple and efficient method for synthesizing pure single stranded RNAs of virtually any structure is described. This in vitro transcription system is based on the unusually specific RNA synthesis by bacteriophage SP6 RNA polymerase which initiates transcription exclusively at
E T Schenborn et al.
Nucleic acids research, 13(17), 6223-6236 (1985-09-11)
The in vitro synthesis of extraneous RNA sequences by SP6 and T7 RNA polymerases from specific DNA templates is described. Transcription of templates prepared by digestion with restriction enzymes that leave 3' protruding ends resulted in the production of significant
M R Green et al.
Cell, 32(3), 681-694 (1983-03-01)
To study the mechanisms of RNA splicing we have synthesized beta-globin mRNA precursors by in vitro transcription using a plasmid in which a human beta-globin gene is fused to an efficient bacteriophage promoter. The structural requirements for accurate splicing of

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