>98% as determined by SDS-PAGE. Purified by Ni-NTA chromatography.
One unit is defined as the amount of the enzyme incorporates 1 nmol of ATP into acid-insoluble product in 1 hour at 37°C.
1X RNA Polymerase Reaction Buffer, supplemented with 3 mM each ATP, UTP, GTP, CTP, and DNA template containing the
T7 RNA Polymerase promoter. Incubate at 37°C.
T7 RNA Polymerase is supplied in 100 mM Tris-HCl (pH 7.9), 20 mM KCl, 1 mM DTT, 1 mM EDTA, 0.1% Triton® X-100 and 50% (v/v)
Stored at -20°C. For optimal storage, aliquot the reaction buffer and DTT reagent into smaller quantities and store at
recommended temperature. For most favorable performance, avoid repeated handling and multiple freeze/thaw cycles.
1. Transcription reaction should be performed under RNase free condition. Use nuclease-free tubes, reagents, and water to avoid
RNase contamination. Also, wear gloves when working with RNA.
2. The volume of T7 RNA Polymerase can be titrated between 1-2 μL in the IVT reaction to optimize your assay.
Standard RNA synthesis procedures:
1. Below reaction mixture should be prepared under room temperature and combined in the following order:
2. Incubate at 37°C for 30 minutes to 2 hours.
3. Above reaction mixture may be scaled up or down proportionately.
|10X RNA Polymerase Reaction Buffer B
|ATP (100 mM)
|UTP (100 mM)
|CTP (100 mM)
|GTP (100 mM)
|100 mM DTT
|T7 RNA Polymerase (200 U/μL)
|RNase inhibitor (optional)
|Total reaction volume