Immobilization and hybridization by single sub-millisecond electric field pulses, for pixel-addressed DNA microarrays

被引:37
作者
Fixe, F
Branz, HM
Louro, N
Chu, V
Prazeres, DMF
Conde, JP
机构
[1] INESC, MN, P-1000029 Lisbon, Portugal
[2] Univ Tecn Lisboa, Ctr Biol & Chem Engn, Inst Super Tecn, P-1049001 Lisbon, Portugal
[3] Natl Renewable Energy Lab, Golden, CO 80401 USA
[4] Univ Tecn Lisboa, Dept Mat Engn, Inst Super Tecn, P-1049001 Lisbon, Portugal
关键词
electric field; DNA; chip; immobilization; hybridization; microarray; rate; voltage; pulsed;
D O I
10.1016/j.bios.2003.12.012
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
Single square voltage pulses applied to buried electrodes result in dramatic rate increases for (1) selective covalent bonding (immobilization) of single-stranded DNA (ssDNA) probes to a functionalized thin film SiO2 surface on a plastic substrate and (2) hybridization of ssDNA to the immobilized probe. DNA immobilization and hybridization times are 100 ns and 10 mus, respectively, about 10(9) times faster than the corresponding passive reactions without electric field. Surface coverage is comparable. Duration, magnitude and slew rate of the voltage pulse are all key factors controlling the rates of ssDNA immobilization and hybridization. With rise times of 4.5 ns, pulses shorter than 1 ms and voltages below 1 V are effective. The ssDNA adsorbed on the surface is reoriented by the rapidly changing electric field. This reduces steric barriers and speeds the immobilization and hybridization reactions. These results open the way for pixel-addressed microarrays driven by silicon microelectronics circuits. (C) 2004 Elsevier B.V. All rights reserved.
引用
收藏
页码:1591 / 1597
页数:7
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