T7 RNA Polymerase，5KU，RT7POL-5KU，Sigma

General description

T7 RNA Polymerase is an RNA polymerase from the T7 bacteriophage that catalyzes the formation of RNA from DNA in the 5′ to 3′ direction. The T7 polymerase also requires a double stranded DNA template and Mg2+ ion as cofactor for the synthesis of RNA. It has a very low error rate.

Application

This product is a recombinant protein expressed in E. coli and induced by IPTG. The MW of T7 RNA Polymerase is 98.9 kDa and theoretical pI of ~6.8.

Features and Benefits

T7 polymerase is extremely promoter-specific, transcribes only DNA downstream of a T7 promoter, and can be used for in vitro mRNA synthesis. Quality tests include:

• Nickase activty: None detected in a 50 µl reaction containing 250 ng of supercoiled pUC19 DNA and at least 30 units of T7 RNA Polymerase incubated for 4 hours at 37°C.
• DNase Activity: None detected in a 50 µl reaction containing 250 ng linear pUC19 DNA and at least 30 units of T7 RNA Polymerase incubated for 16 hours at 37°C.
• RNase Activity: None detected in a 50 µl reaction containing 1.25-2.5 ng tRNA and at least 30 units of T7 RNA Polymerase is incubated for 4h at 37°C.

Physical form

The product is supplied as an aqueous solution containing 50mM Tris-HCl, 100mM NaCl, 20mM β-ME, 1mM EDTA, 0.02% (w/v) Tween 20 and 50% glycerol, pH 7.9 at 25 °C.

Other Notes

1 Unit of T7 RNA polymerase is defined as the amount of enzyme that synthesizes 1ug of mRNA per hour in a 30-minute reaction containing 25μg/ml linearized plasmid, 3mM of each NTP and 18mM MgCl2, at 37°C, pH 7.7