Preparation of linear plasmid DNA for in vitro transcription reaction

Linearised pDNA is currently the starting point of In-Vitro-Transcription processes to synthesize mRNA. Large scale purification protocols for manufacturing of pDNA used for Gene Therapy applications typically include two chromatography steps. The first step captures both linear, open circular and supercoiled pDNA species. The polishing step enriches supercoiled pDNA, while discarding other isoforms. We describe a single-step-capture strategy to maximize the recovery of pDNA for further linearization.