Dielectrophoretic separation of E. coli bacteria from a mixture containing blood cells followed by electronic lysis was performed on a microfabricated bioelectronic chip by applying a series of high voltage pulses. Bacteria were isolated from human blood cells using dielectrophoresis to direct the prokaryotic cells to 25 microlocations directly above individually-addressable platinum microelectrodes on a chip. The surface of the chip, including the platinum electrodes and the rest of the non-metallized area, was coated with an agarose permeation layer to prevent the direct contact of cells with the electrode and also to minimize the non-specific adhesion of cells and lysate to the chip surface. The platinum electrodes were 80μm in diameter and had a center-to-center spacing of 200μm. Analysis of the electronically-lysed bacterial mixture showed that it contained a spectrum of nucleic acids including RNA, plasmid DNA and genomic DNA. The lysate was then further analysed by electronically-enhanced nucleic acid hybridization on different bioelectronic chips made for specific DNA and RNA hybridization analyses. Dielectrophoretic cell separation followed by electronic lysis has significant potential as a sample preparation process for chip-based electronic-hybridization assays in an integrated DNA/RNA analysis system.