Description
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.
Application Applications include:
• Synthesis of labeled and unlabeled RNA transcripts
• RNase protection assays
• Generate RNA transcripts for in vitro translation
 
Biochem/physiol Actions 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. The enzyme is ideal for in vitro transcription of RNA transcripts, antisense RNA （from reversed cloned DNA insert）, and biologically active mRNA that can be precisely spliced. RNA transcripts are a choice in the creation of RNA:DNA hybridization probes because of their stability. Sp6 polymerase exhibits 30% more activity at 40 °C than at 37 °C.
Unit Definition One unit will catalyze the incorporation of 1 nmol of rNTP into acid-precipitable material in 60 min at 37 °C.
Physical form Solution in 100 mM NaCl, 50 mM Tris-HCl, pH 7.9, 0.1 mM EDTA, 0.1% Triton X-100, 1 mM DTT, 50% （v/v） glycerol

Properties
form buffered aqueous solution
concentration 10,000-50,000 units/mL
foreign activity DNase, RNase, protease, none detected
  nonspecific endonuclease, absence of activity （after 20 hour incubation）
shipped in dry ice
storage temp. −20°C

Safety
WGK Germany 1

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