Label-Free Identification of Single Mononucleotides by Nanoscale Electrophoresis,Small

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Label-Free Identification of Single Mononucleotides by Nanoscale Electrophoresis
Small ( IF 13.0 ) Pub Date : 2021-09-23 , DOI: 10.1002/smll.202102567
Junseo Choi _{1,

2} , Zheng Jia _{1,

2} , Ramin Riahipour _{1,

2} , Collin J McKinney _{2,

3} , Charuni A Amarasekara _{2,

4} , Kumuditha M Weerakoon-Ratnayake _{2,

4} , Steven A Soper _{2,

4,

5,

6} , Sunggook Park _{1,

2}

Affiliation

Nanoscale electrophoresis allows for unique separations of single molecules, such as DNA/RNA nucleobases, and thus has the potential to be used as single molecular sensors for exonuclease sequencing. For this to be envisioned, label-free detection of the nucleotides to determine their electrophoretic mobility (i.e., time-of-flight, TOF) for highly accurate identification must be realized. Here, for the first time a novel nanosensor is shown that allows discriminating four 2-deoxyribonucleoside 5'-monophosphates, dNMPs, molecules in a label-free manner by nanoscale electrophoresis. This is made possible by positioning two sub-10 nm in-plane pores at both ends of a nanochannel column used for nanoscale electrophoresis and measuring the longitudinal transient current during translocation of the molecules. The dual nanopore TOF sensor with 0.5, 1, and 5 µm long nanochannel column lengths discriminates different dNMPs with a mean accuracy of 55, 66, and 94%, respectively. This nanosensor format can broadly be applicable to label-free detection and discrimination of other single molecules, vesicles, and particles by changing the dimensions of the nanochannel column and in-plane nanopores and integrating different pre- and postprocessing units to the nanosensor. This is simple to accomplish because the nanosensor is contained within a fluidic network made in plastic via replication.

中文翻译：

通过纳米级电泳无标记鉴定单个单核苷酸

纳米级电泳允许对单分子进行独特的分离，例如 DNA/RNA 核碱基，因此有可能用作外切核酸酶测序的单分子传感器。为此，必须实现核苷酸的无标记检测以确定它们的电泳迁移率（即，飞行时间，TOF），以实现高度准确的识别。在这里，首次展示了一种新型纳米传感器，该传感器允许通过纳米级电泳以无标记方式区分四种 2-脱氧核糖核苷 5'-单磷酸、dNMP、分子。这是通过在用于纳米级电泳的纳米通道柱的两端放置两个亚 10 nm 平面内孔并测量分子易位期间的纵向瞬态电流来实现的。双纳米孔 TOF 传感器具有 0.5、1、和 5 µm 长的纳米通道柱长度区分不同的 dNMP，平均准确度分别为 55%、66% 和 94%。通过改变纳米通道柱和平面内纳米孔的尺寸并将不同的预处理和后处理单元集成到纳米传感器中，这种纳米传感器格式可广泛应用于其他单分子、囊泡和颗粒的无标记检测和区分。这很容易实现，因为纳米传感器包含在通过复制由塑料制成的流体网络中。通过改变纳米通道柱和平面内纳米孔的尺寸并将不同的预处理和后处理单元集成到纳米传感器中来实现颗粒和颗粒。这很容易实现，因为纳米传感器包含在通过复制由塑料制成的流体网络中。通过改变纳米通道柱和平面内纳米孔的尺寸并将不同的预处理和后处理单元集成到纳米传感器中来实现颗粒和颗粒。这很容易实现，因为纳米传感器包含在通过复制由塑料制成的流体网络中。

更新日期：2021-10-21

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