5.6.2.5. Synthesizing full-length viral RNA

The plasmid can be used as a template for in vitro transcription using T7 RNA polymerase.

1. Linearize plasmid downstream of the 3’ poly-A sequence with PmeI restriction endonuclease.
2. Purify the linearized templates with a Qiagen plasmid purification kit.
3. Synthesize capped in vitro RNA transcripts using “mMESSAGEmMAchine” T7 kit (Ambion).