The need for large quantities of purified plasmid DNA has increased as the applications of DNA vaccines continue to expand. This chapter describes a simple, scaleable procedure based on the fed-batch cultivation of various Escherichia coli clones, which can be easily implemented and scaled-up to large bioreactors. Although some clones may require minor modifications to the feeding strategy, in general, this procedure, implemented as described, is likely to support the production of milligram to gram quantities of plasmid DNA.