Detecting Single Molecule Deoxyribonucleic Acid in a Cell Using a Three-Dimensionally Integrated Nanopore

Small Methods. 2021 Sep;5(9):e2100542. doi: 10.1002/smtd.202100542. Epub 2021 Aug 15.

Abstract

Amplification-free genome analysis can revolutionize biology and medicine by uncovering genetic variations among individuals. Here, the authors report on a 3D-integrated nanopore for electrolysis to in situ detection of single-molecule DNA in a cell by ionic current measurements. It consists of a SiO2 multipore sheet and a SiNx nanopore membrane stacked vertically on a Si wafer. Single cell lysis is demonstrated by 106 V m-1 -level electrostatic field focused at the multinanopore. The intracellular molecules are then directly detected as they move through a sensing zone, wherein the authors find telegraphic current signatures reflecting folding degrees of freedom of the millimeter-long polynucleotides threaded through the SiNx nanopore. The present device concept may enable on-chip single-molecule sequencing to multi-omics analyses at a single-cell level.

Keywords: cell lysis; deoxyribonucleic acid; electrophoresis; nanoelectromechanical systems; nanopores.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biosensing Techniques
  • DNA / analysis*
  • Humans
  • Nanopores
  • Silicon Dioxide / chemistry
  • Single Molecule Imaging / instrumentation*
  • Single Molecule Imaging / methods
  • Static Electricity

Substances

  • Silicon Dioxide
  • DNA