Patent ID: 9487788
Filing Date: 2016-11-08
CPC Classification: C12N

Claim Text:
1. A method for increasing production yield of a covalently closed, super-coiled plasmid DNA vector that is non-repetitive and does not contain direct repeats or long terminal repeats, comprising the steps of: a. transforming a plasmid DNA vector that is non-repetitive and does not contain direct repeats or long terminal repeats into bacterial cells rendered competent for transformation; and b. growing resultant transformed bacterial cells at a reduced temperature during colony isolation and colony amplification steps in order to create a cryopreserved seed stock for plasmid DNA vector production, the reduced temperature being approximately 20-33° C.; whereby growth at said reduced temperature prior to cryopreservation improves said plasmid DNA vector yield in subsequent shake flask and or fermentation culture above yield that is achieved when growing the transformed bacterial cells at a temperature approximately 33-42° C. during colony isolation and colony amplification steps.