Periodic DNA nanotemplates synthesized by rolling circle amplification

Nano Lett. 2005 Apr;5(4):719-22. doi: 10.1021/nl050155a.

Abstract

Rolling circle amplification (RCA) is an elegant biochemical method by which long single-stranded DNA molecules with a repeating sequence motif can be readily synthesized. In RCA, small circular single-stranded oligonucleotides serve as templates for the polymerization of the complementary strand. A DNA polymerase with an efficient strand displacement activity can copy the circular template without stopping. This results in a long DNA strand with periodic sequence. We here demonstrate that this method, using DNA recognition and biotin-streptavidin binding, provides a simple procedure for DNA-directed nanoscale organization of matter. As an example, a 74 nucleotide (nt) long circular DNA molecule is amplified into a sequence-periodic single strand with a length up to several micrometers. Hybridization of this long periodic DNA template to the biotinylated complement of the sequence motif results in a long DNA duplex with a periodic arrangement of biotin binding sites. On this duplex, streptavidin-coated particles can be organized into one-dimensional arrays. The resulting DNA constructs are characterized by gel electrophoresis and atomic force microscopy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biotin / chemistry
  • Biotinylation
  • DNA Replication*
  • DNA, Circular / chemistry*
  • DNA, Single-Stranded / biosynthesis*
  • DNA-Directed DNA Polymerase / chemistry
  • Nanostructures / chemistry*
  • Nucleic Acid Hybridization
  • Oligodeoxyribonucleotides / chemistry
  • Streptavidin / chemistry
  • Templates, Genetic*

Substances

  • DNA, Circular
  • DNA, Single-Stranded
  • Oligodeoxyribonucleotides
  • Biotin
  • Streptavidin
  • DNA-Directed DNA Polymerase