High Yield T7 RNA Synthesis Kit (≥2000 nt)

Storage Conditions

All components should be stored at -20°C

Unit Definition

One unit is defined as the amount of enzyme that will incorporate 1 nmol ATP into acid-insoluble material in a total reaction volume of 50 µl in 1 hour at 37°C in 1X RNA Polymerase Reaction Buffer.

Quality Control

Exonuclease Activity: Incubation of a 40 µl reaction containing a minimum of 40 units of T7 RNA Polymerase with 2.5 μg λ-Hind III digest DNA for 16 h at 37℃ results in no detectable degradation as determined.

Endonuclease Activity: Incubation of a 50 µl reaction containing a minimum of 50 units of T7 RNA Polymerase with 1 μg λDNA for 16 h at 37℃ results in no detectable degradation as determined.

RNase Activity: Incubation of a 40 µl reaction containing a minimum of 40 units of T7 RNA Polymerase with 1 μg MS2 RNA for 2 h at 37 ℃ results in no detectable degradation as determined.

DNase Activity: Incubation of a 40 µl reaction containing a minimum of 40 units of T7 RNA Polymerase with 4 μg pUC19 DNA for 16 h at 37℃ results in no detectable degradation as determined.

Protocol

1. Assemble the reaction at room temperature in the following order: