Atomic force microscopy imaging of double stranded dna and rna

Yuri L. Lyubchenko, Bertram L. Jacobs, Lyuda S. Shlyakhtenko, Rodney E. Harrington, Alexander A. Gall, Patrick I. Oden, Stuart M. Lindsay

Research output: Contribution to journalArticle

141 Scopus citations

Abstract

A procedure for imaging long DNA and double stranded RNA (dsRNA) molecules using Atomic Force Microscopy (AFM) is described. Stable binding of double stranded DNA molecules to the flat mica surface is achieved by chemical modification of freshly cleaved mica under mild conditions with 3-aminopropyltriethoxy silane. We have obtained striking images of intact lambda DNA, Hind III restriction fragments of lambda DNA and dsRNA from reovirus. These images are stable under repeated scanning and measured contour lengths are accurate to within a few percent. This procedure leads to strong DNA attachment, allowing imaging under water. The widths of the DNA images lie in the range of 20 to 80nm for data obtained in air with commercially available probes. The work demonstrates that AFM is now a routine tool for simple measurements such as a length distribution. Improvement of substrate and sample preparation methods are needed to achieve yet higher resolution.

Original languageEnglish (US)
Pages (from-to)589-606
Number of pages18
JournalJournal of Biomolecular Structure and Dynamics
Volume10
Issue number3
DOIs
StatePublished - Dec 1992

ASJC Scopus subject areas

  • Structural Biology
  • Molecular Biology

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    Lyubchenko, Y. L., Jacobs, B. L., Shlyakhtenko, L. S., Harrington, R. E., Gall, A. A., Oden, P. I., & Lindsay, S. M. (1992). Atomic force microscopy imaging of double stranded dna and rna. Journal of Biomolecular Structure and Dynamics, 10(3), 589-606. https://doi.org/10.1080/07391102.1992.10508670