Ying Liming, White Samuel S, Bruckbauer Andreas, Meadows Lisa, Korchev Yuri E, Klenerman David
Department of Chemistry, University of Cambridge, Cambridge, CB2 1EW, United Kingdom.
Biophys J. 2004 Feb;86(2):1018-27. doi: 10.1016/S0006-3495(04)74177-6.
The study of the properties of DNA under high electric fields is of both fundamental and practical interest. We have exploited the high electric fields produced locally in the tip of a nanopipette to probe the motion of double- and single-stranded 40-mer DNA, a 1-kb single-stranded DNA, and a single-nucleotide triphosphate (dCTP) just inside and outside the pipette tip at different frequencies and amplitudes of applied voltages. We used dual laser excitation and dual color detection to simultaneously follow two fluorophore-labeled DNA sequences with millisecond time resolution, significantly faster than studies to date. A strong trapping effect was observed during the negative half cycle for all DNA samples and also the dCTP. This effect was maximum below 1 Hz and decreased with higher frequency. We assign this trapping to strong dielectrophoresis due to the high electric field and electric field gradient in the pipette tip. Dielectrophoresis in electrodeless tapered nanostructures has potential applications for controlled mixing and manipulation of short lengths of DNA and other biomolecules, opening new possibilities in miniaturized biological analysis.
对高电场下DNA特性的研究具有基础和实际意义。我们利用纳米移液器尖端局部产生的高电场,在不同频率和施加电压幅度下,探测移液器尖端内外双链和单链40聚体DNA、1千碱基单链DNA以及单核苷酸三磷酸(dCTP)的运动。我们使用双激光激发和双色检测,以毫秒级时间分辨率同时跟踪两个荧光团标记的DNA序列,比迄今为止的研究速度快得多。在所有DNA样品以及dCTP的负半周期期间都观察到了强烈的捕获效应。这种效应在低于1赫兹时最大,并随着频率升高而降低。我们将这种捕获归因于移液器尖端的高电场和电场梯度导致的强介电泳。无电极锥形纳米结构中的介电泳在短长度DNA和其他生物分子的受控混合与操纵方面具有潜在应用,为小型化生物分析开辟了新的可能性。