Motion and Enzymatic Degradation of DNA in the Atomic Force Microscope

Biophys J. 1994 Dec;67(6):2454-9. doi: 10.1016/S0006-3495(94)80733-7.

Abstract

The dynamics and enzymatic degradation of single DNA molecules can now be observed with the atomic force microscope. A combination of two advances has made this possible. Tapping in fluid has reduced lateral forces, which permits the imaging of loosely adsorbed molecules; and the presence of nickel ions appears to form a relatively stable bridge between the negatively charged mica and the negatively charged DNA phosphate backbone. Continuous imaging shows DNA motion and the process of DNA degradation by the nuclease DNase I. It is possible to see DNase degradation of both loosely adsorbed and tightly adsorbed DNA molecules. This method gives images in aqueous buffer of bare, uncoated DNA molecules with lengths of only a few hundred base pairs, or approximately 100 nm in length.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Adsorption
  • Biophysical Phenomena
  • Biophysics
  • Buffers
  • DNA / chemistry*
  • DNA / ultrastructure*
  • Deoxyribonuclease I
  • Microscopy, Atomic Force / instrumentation
  • Microscopy, Atomic Force / methods*
  • Motion
  • Nickel
  • Water

Substances

  • Buffers
  • Water
  • Nickel
  • DNA
  • Deoxyribonuclease I