SP6 RNA Polymerase is a DNA-dependent RNA polymerase with a high sequence specificity for SP6 promoter sequences. See Figure 1 for gel analysis using SP6 RNA polymerase. SP6 RNA polymerase synthesizes RNA 5' to 3' and incorporates 35S, 32P and 33P ribonucleotides. This enzyme is isolated from an overproducing recombinant E. coli clone.
Unit Definition: One unit will catalyze the incorporation of 1 nmole of nucleoside triphosphate into an acid insoluble form in 60 minutes at 37°C.
Purity: Determined to be 99% pure by SDS-polyacrylamide gel electrophoresis analysis. The absence of contaminating DNase and RNase activity is confirmed by the integrity of RNA transcripts produced by SP6 polymerase when analyzed on a denaturing polyacrylamide gel.
Reaction Conditions: 40 mM Tris-HCl (pH 8.0), 8 mM MgCl2, 50 mM NaCl, 2 mM spermidine, 30 mM DTT, 400 µM of each rNTP, 1 µg DNA template and 20 units enzyme in a 25-µl volume. Incubate 60 minutes at 37°C.
For Research Use Only. Not for use in diagnostic procedures