Electric field directed nucleic acid hybridization on microchips.
Selection and adjustment of proper physical parameters enables rapid DNA transport, site selective concentration, and accelerated hybridization reactions to be carried out on active microelectronic arrays. These physical parameters include DC current, voltage, solution conductivity and buffer species. Generally, at any given current and voltage level, the transport or mobility of DNA is inversely proportional to electrolyte or buffer conductivity. However, only a subset of buffer species produce both rapid transport, site specific concentration and accelerated hybridization. These buffers include zwitterionic and low conductivity species such as: d- and l-histidine; 1- and 3-methylhistidines; carnosine; imidazole; pyridine; and collidine. In contrast, buffers such as glycine, beta-alanine and gamma-amino-butyric acid (GABA) produce rapid transport and site selective concentration but do not facilitate hybridization. Our results suggest that the ability of these buffers (histidine, etc.) to facilitate hybridization appears linked to their ability to provide electric field concentration of DNA; to buffer acidic conditions present at the anode; and in this process acquire a net positive charge which then shields or diminishes repulsion between the DNA strands, thus promoting hybridization.
Edman
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CF
Nanogen, 10398 Pacific Center Court, San Diego, CA 92121, USA.
Raymond
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DE
Wu
D J
DJ
Tu
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E
Sosnowski
R G
RG
Butler
W F
WF
Nerenberg
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Heller
M J
MJ
D002021:Buffers; D004247:DNA / Q000737:chemistry; D004574:Electromagnetic Fields; D004581:Electronics / Q000295:instrumentation*; D008832:Microchemistry / Q000295:instrumentation*; D008904:Miniaturization; D009693:Nucleic Acid Hybridization; D012666:Semiconductors
D002021:Buffers; D004247:DNA