22.214.171.124. 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
2. Purify the linearized templates with a Qiagen plasmid purification kit.
3. Synthesize capped in vitro RNA transcripts using “mMESSAGEmMAchine” T7 kit (Ambion).