Plasmid deoxyribonucleic acid (pDNA) is a promising therapeutic in gene therapy and as a DNA vaccine, but it is susceptible to degradation by nucleases in the extracellular environment. In this study, we report on the enhanced enzymatic degradation resistance of pDNA, when stored in a hydrated buffered ionic liquid (BIL) based on choline dihydrogen phosphate (CDHP). The stability of pDNA stored in buffered CDHP (bCDHP) in the presence of Turbo DNase was studied using agarose gel electrophoresis, which showed a prolonged shelf life at room temperature over a period of 28 days in bCDHP compared to within 10 minutes for phosphate buffered saline (PBS). In addition, the biological activity of pDNA was maintained under such conditions, as demonstrated by the expression of yellow fluorescent protein (YFP).