Extraction of nucleic acid in Escherichia coli and Saccharomyces cerevisiae using high electric field pulse application

Abstract

AbstractAn electroporation that formed the pores on cell membrane after application of high electric field pulses has been used the transformation and the sterilization of cell. In this paper, we reported the extraction method of nucleic acid in Escherichia coli and Saccharomyces cerevisiae using high electric field pulse application. The sample was 1.0 × 10–3 mol/L NaCl aqueous solution contained E. coli and S. cerevisiae. High voltage pulse is applied to the sample at room temperature. These results were shown as the following: (1) When a pulsed electric field of 4 ∼ 13 kV/mm is applied to E. coli and S. cerevisiae, the estimated amount of nucleic acid increases with the increases of strength of applied pulsed electric field. (2) In the case of E. coli, the 2–3 kbp band was confirmed at an electric field of 5 kV/mm or more, and the 10 kbp band was confirmed at an electric field of 8 kV/mm or more. (3) In the case of S. cerevisiae, the 2–3 kbp band was confirmed at an electric field of 9 kV/mm or more. the 10 kbp band could not be confirmed at 13 kV/mm or less.