FAQ: Can an uncut plasmid be used as a template for the HiScribe™ T7 mRNA Kit with CleanCap® Reagent AG?

No. T7 RNA Polymerase is an extremely processive enzyme and will continue to transcribe around a circular template multiple times without disassociating. The plasmid can be linearized with restriction enzymes that leave a blunt or 5′ overhang downstream of the DNA of interest resulting in run-off transcription. See the product manual for DNA template generation.