Preparation and hybridization analysis of DNA/RNA from E. coli on microfabricated bioelectronic chips.
Escherichia coli were separated from a mixture containing human blood cells by means of dielectrophoresis and then subjected to electronic lysis followed by proteolytic digestion on a single microfabricated bioelectronic chip. An alternating current electric field was used to direct the bacteria to 25 microlocations above individually addressable platinum microelectrodes. The platinum electrodes were 80 microns in diameter and had center-to-center spacings of 200 microns. After the isolation, the bacteria were lysed by a series of high-voltage pulses. The lysate contained a spectrum of nucleic acids including RNA, plasmid DNA, and genomic DNA. The lysate was further examined by electronically enhanced hybridization on separate bioelectronic chips. Dielectrophoretic separation of cells followed by electronic lysis and digestion on an electronically active chip may have potential as a sample preparation process for chip-based hybridization assays in an integrated DNA/RNA analysis system.
Cheng
J
J
Nanogen, Inc., San Diego, CA 92121, USA. jcheng@nanogen.com
Sheldon
E L
EL
Wu
L
L
Uribe
A
A
Gerrue
L O
LO
Carrino
J
J
Heller
M J
MJ
O'Connell
J P
JP
D015374:Biosensing Techniques; D004269:DNA, Bacterial / Q000302:isolation & purification*; D004581:Electronics / Q000295:instrumentation; D004586:Electrophoresis / Q000379:methods; D004926:Escherichia coli / Q000235:genetics*; D006801:Humans; D008839:Microelectrodes; D008904:Miniaturization; D009693:Nucleic Acid Hybridization / Q000379:methods*; D012329:RNA, Bacterial / Q000302:isolation & purification*
D004269:DNA, Bacterial; D012329:RNA, Bacterial