T7, RNA Polymerases produce defined RNA by in vitro transcription of DNA cloned into a plasmid or other vector downstream from their respective promoter.
Note that these are supplied as enzyme only1.
Notes
- A buffer package containing 5ml of 5X Transcription Buffer and 2.5ml of 100mM DTT should be purchased separately.
- For synthesis of large amounts of RNA, use the AmpliScribe™ High Yield Transcription Kits or for synthesis of large amounts of 5'-capped RNA use the AmpliCap™ High Yield Message Maker Kits.
- For synthesis of high yields of RNA that is completely resistant to RNase A digestion, use the DuraScribe® T7 Transcription Kit.
Unit Definition
One unit catalyses the incorporation of 1 nmole of a labelled ribonucleoside triphosphate into RNA in 1 hour at 37°C in 40mM Tris-HCl, pH 7.5, 6mM MgCl2, 10mM NaCl, 2mM spermidine, 10mM DTT, and 0.5mM each NTP using a DNA template with the appropriate T7, SP6, or T3 promoter.
Storage Buffer
50% glycerol containing 50mM Tris-HCl, pH 7.5, 0.1M NaCl, 1.0mM DTT, 0.1mM EDTA, and 0.1% Triton X-100.
5X Transcription Buffer
0.2M Tris-HCl, pH 7.5, 50 mM NaCl, 30mM MgCl2, and 10mM spermidine.
Quality Control
These RNA Polymerases are tested in transcription and are free of detectable exo- and endonuclease and RNase activities, and E. coli RNA polymerase.
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