T7 RNA polymerase is a highly specific DNA dependent RNA polymerase originated from T7 bacteriophage. One of the most potent tools used for the study of RNA dynamics.
(SKU: RN003)
€50.00
T7 RNA polymerase is a highly specific DNA dependent RNA polymerase originated from T7 bacteriophage. One of the most potent tools used for the study of RNA dynamics.
(SKU: RN003)
Temporarily discontinued!
T7 RNA polymerase is a highly specific DNA dependent RNA polymerase originated from T7 bacteriophage. One of the most potent tools used for the study of RNA dynamics.
Applications
Transcription Guidelines
200U T7 Polymerase | 2μl |
10x T7 Buffer | 2μl |
DNA substrate* | 1 μg** |
RNTPs mix (40 mM) | 2 μl |
RNase inhibitor | 20 u |
Sterile ultrapure water | Up to 20 μl |
Incubate for 120 min at 37oC |
*PCR products, annealed oligos or digested plasmid containing T7 promoter Sequence: TAATACGACTCACTATA, ** 0,1 to 2 μg of template DNA have been successfully tested yielding on average 10-40 times of the RNA amount eg. 15 μg of RNA were produced from 250 ng of pre annealed DNA oligos containing T7 RNA promoter.
Setup guidelines:
Make sure that buffer has been thawed completely (no white precipitant due to DTT precipitation is visible). Thaw well at room temperature. Add reactants with the following order : water, buffer (mix well) , RNAse inhibitor, template (mix well), RNTPs, T7 RNA polymerase.
Provided with 10 X T7 RNA pol buffer: 400 mM Tris-HCl (pH 7.9 @ 25°C) 80 mM MgCl2 10mM DTT 20mM spermidine.
Inactivation: Heat inactivation at 70oC for 20 minutes or by the addition of EDTA at a final of 5 mM (not recommended when EDTA may inhibit downstream applications)
Storage conditions:
Non-specific nuclease activity: 200 units of T7 RNA polymerase do not produce any unspecific cleavage products after 6 hrs incubation with neither 1 μg of Lambda DNA nor pBR322 at 37oC .
No RNASE activity: 200 units of T7 RNA polymerase do not produce any degradation after 6 hrs incubation with total RNA from HeLa cells.
Concentration | 100u/μl |
---|---|
Quantity | 5000 U |
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