SP6 RNA Polymerase
High concentration available Recombinant enzyme Thermal inactivation at 70°C in 10 min
SP6 RNA polymerase is DNA-dependent RNA polymerase catalyzes the 5'→3' synthesis of RNA on either single-stranded or double-stranded DNA under control of the SP6 promoter
Thermo Scientific Bacteriophage SP6 RNA polymerase is a DNA-dependent RNA polymerase with strict specificity for its respective double-stranded promoters. It catalyzes the 5'→3' synthesis of RNA on either single-stranded DNA or double-stranded DNA downstream from its promoter and incorporates modified nucleotides.
- Incorporates modified nucleotides (e.g., aminoallyl-, biotin-, fluorescein-, digoxigenin-labeled nucleotides)
- Synthesis of unlabeled and labeled RNA that can be used:
- For hybridization (see Reference 1), in vitro RNA translation (see Reference 2)
- As aRNA (see Reference 3), siRNA (see Reference 4), substrate in RNase protection assays (see Reference 5), template for genomic DNA sequencing (see Reference 6)
- In studies of RNA secondary structure and RNA-protein interactions (see Reference 7), RNA splicing (see Reference 8)
Consensus promoter sequences:
The position in bold (+1) indicates the first nucleotide incorporated into RNA during transcription. Only bases at this position through +3 are critical for transcription, and they must be a G and a purine base, respectively (see Reference 9).
|5× Transcription Buffer||200 mM Tris-HCl (pH 7.9 at 25°C), 30 mM MgCl2, 50 mM DTT, 50 mM NaCl, 10 mM spermidine.|
|Definition of Activity Unit||One unit of the enzyme incorporates 1 nmol of AMP into a polynucleotide fraction (adsorbed on DE-81) in 60 minutes at 37°C. Enzyme activity is assayed in the following mixture: 40 mM Tris-HCl (pH 8.0), 6 mM MgCl2, 10 mM DTT, 2 mM spermidine, 0.5 mM of each NTP, 0.6 MBq/mL [3H]-ATP, 20 µg/mL plasmid DNA containing the appropriate promoter sequences.|
|Inactivation||Inactivated by heating at 70°C for 10 min or by addition of EDTA.|
|Inhibition||Inhibitors: metal chelators, enzyme activity is reduced by 50% at NaCl or KCl concentration above 150 mM. Greater than 50% reduction in enzyme activity with ammonium sulphate.|
|Molecular Weight||99 kDa monomer|
|Source||E. coli cells with a cloned gene encoding SP6 RNA polymerase.|
|Storage Buffer||Polymerase is supplied in 50 mM Tris-HCl (pH 8.0), 150 mM NaCl, 5 mM DTT, 0.1 mg/mL BSA, 0.03% (v/v) ELUGENT Detergent, 50% (v/v) glycerol.|
|Storage Condition||-20 C|
Produkt zatím nikdo nehodnotil, buďte první!