DNA characterization with ion beam-sculpted silicon nitride nanopores.
DNA characterization with ion beam-sculpted silicon nitride nanopores.
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DOI：
10.1007/978-1-61779-773-6_5
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发表时间：
2012
期刊：
影响因子：
--
通讯作者：
Li, Jiali
中科院分区：
文献类型：
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作者：
Rollings, Ryan C;McNabb, David S;Li, Jiali
固态纳米孔正在成为强大的单分子电子测量装置和限制生物分子进行进一步分析的平台。第一个用于检测单个 DNA 分子的氮化硅纳米孔是使用离子束雕刻 (IBS) 制造的，这种方法使用宽幅低能离子束来创建尺寸范围为 2 至 20 nm 的纳米孔。在本章中，我们讨论 IBS 雕刻纳米孔的制造、表征和使用，以及有效使用 pClamp 和 MATLAB 软件套件进行数据采集和分析。制造部分将涵盖重复性和孔径限制。表征讨论重点关注通过低分辨率和高分辨率透射电子显微镜 (TEM)、电子能量损失光谱 (EELS) 和能量过滤 TEM (EFTEM) 测量的几何特性。易位实验部分重点介绍如何使用纳米孔实验者常用的工具来确定某个孔是否对实验有用或是否应该放弃。将介绍一种使用 Clampex 事件驱动模式和双通道记录获取数据的内存有效方法，然后介绍一种使用 MATLAB 软件轻松实现的多阈值事件检测和分类方法。
Solid state nanopores are emerging as robust single molecule electronic measurement devices and as platforms for confining biomolecules for further analysis. The first silicon nitride nanopore to detect individual DNA molecules were fabricated using ion beam sculpting (IBS), a method that uses broad, low energy ion beams to create nanopores with dimensions ranging from 2 to 20 nm. In this chapter, we discuss the fabrication, characterization, and use of IBS sculpted nanopores as well as efficient uses of pClamp and MATLAB software suites for data acquisition and analysis. The fabrication section will cover the repeatability and the pore size limits. The characterization discussion focuses on the geometric properties as measured by low and high resolution transmission electron microscopy (TEM), electron energy loss spectroscopy (EELS), and energy filtered TEM (EFTEM). The section on translocation experiments focuses on how to use tools commonly available to the nanopore experimenter to determine whether a pore will be useful for experimentation or if it should be abandoned. A memory efficient method of taking data using Clampex’s event-driven mode and dual channel recording will be presented, followed by an easy to implement multi-threshold event detection and classification method using MATLAB software.