02/27/2025
High throughput purification of plasmid DNA and in vitro transcribed mRNA using a high-speed centrifuge
Pascal Rowart, et al. , Eppendorf Application Technologies S.A.
In this Application Note, we show how the Centrifuge CR22N can be further used to purify plasmid DNA and PCR products, which serves as a first step in the mRNA production workflow of in vitro transcription.
To this end, we harvested a large bacterial culture containing a plasmid DNA, grown in the InnovaR S44i Shaker. Rotor R9A2 was used to pellet 3 L of bacterial culture.
The DNA purification of the entire bacterial pellet from one 1.5 L bottle was performed using a combination of the Rotor R15A and the Rotor R22A4. Finally, we show that high-quality transcription processes can produce the mRNA by in vitro transcription.
The entire procedure followed the protocol and recommendations of the research papers by Rossi et al.