Label-free DNA sequencing using Millikan detection.
Label-free DNA sequencing using Millikan detection.
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DOI：
10.1016/j.ab.2015.06.036
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发表时间：
2015-10-15
影响因子：
2.9
通讯作者：
Farinas J
中科院分区：
文献类型：
--
作者：
Dettloff R;Leiske D;Chow A;Farinas J
基于密立根油滴实验的原理，建立了一种无标记DNA测序方法。当聚合酶将核苷酸添加到附着于珠粒的DNA模板时，这种合成测序方法感测珠粒电荷的增加。取决于珠粒上核苷酸电荷数量的电力与相反的流体动力学阻力和恢复系链力之间的平衡导致珠粒速度是附接至珠粒的核苷酸数量的函数。使用暗场显微镜测量通过聚合物拴系到微流体通道并经受振荡电场的珠的速度，并用于确定在每个合成测序循环期间掺入多少核苷酸。在DNA聚合期间可靠地检测到珠速度增加约1%，从而允许对短DNA模板进行测序。该方法可能导致低成本，高通量测序平台，可以在医疗应用中实现常规测序。
A label-free method for DNA sequencing based on the principle of the Millikan oil drop experiment was developed. This sequencing-by-synthesis approach sensed increases in bead charge as nucleotides were added by a polymerase to DNA templates attached to beads. The balance between an electrical force, which was dependent on the number of nucleotide charges on a bead, and opposing hydrodynamic drag and restoring tether forces resulted in a bead velocity that was a function of the number of nucleotides attached to the bead. The velocity of beads tethered via a polymer to a microfluidic channel and subjected to an oscillating electric field was measured using dark-field microscopy and used to determine how many nucleotides were incorporated during each sequencing-by-synthesis cycle. Increases in bead velocity of ~ 1% were reliably detected during DNA polymerization allowing for sequencing of short DNA templates. The method could lead to a low-cost, high-throughput sequencing platform that could enable routine sequencing in medical applications.