Efficient procedure of preparation and properties of long uniform G4-DNA nanowires

Natalia Borovok, Tatiana Molotsky, Jamal Ghabboun, Danny Porath, Alexander Kotlyar

Research output: Contribution to journalArticlepeer-review

Abstract

Here we describe a novel and efficient procedure for preparation of long uniform G4-DNA wires. The procedure includes (i) enzymatic synthesis of double-stranded DNA molecules consisting of long (up to 10,000 bases), continuous G strands and chains of complementary (dC)20-oligonucleotides, poly(dG)-n(dC)20; (ii) size exclusion HPLC separation of the G strands from the (dC)20 oligonucleotides in 0.1 M NaOH; and (iii) folding of the purified G strands into G4-DNA structures by lowering the pH to 7.0. We show by atomic force microscopy (AFM) that the preparation procedure yielded G4-DNA wires with a uniform morphology and a narrow length distribution. The correlation between the total amount of nucleotides in the G strands and the contour length of the G4-DNA molecules estimated by AFM suggests monomolecular folding of the G strands into quadruplex structures. The folding takes place either in the presence or in the absence of stabilizing ions (K+ or Na+). The addition of these cations leads to a dramatic change in the circular dichroism spectrum of the G4-DNA.

Original languageEnglish
Pages (from-to)71-78
Number of pages8
JournalAnalytical Biochemistry
Volume374
Issue number1
DOIs
StatePublished - 1 Mar 2008

Keywords

  • AFM
  • CD spectroscopy
  • Enzymatic synthesis
  • Folding of poly(dG) strands
  • G4-DNA nanowires

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