Hybridization and enzymatic extension of Au nanoparticle-bound oligonucleotides

Sheila R. Nicewarner Peña, Surabhi Raina, Glenn P. Goodrich, Nina V. Fedoroff, Christine D. Keating

Research output: Contribution to journalArticlepeer-review

135 Scopus citations

Abstract

We have investigated the impact of steric effects on the hybridization and enzymatic extension of oligonucleotides bound to 12-nm colloidal Au particles. In these experiments, a nanoparticle-bound 12-mer sequence is hybridized either to its solution phase 12-mer complement or to an 88-mer template sequence. The particle-bound oligonucleotide serves as a primer for enzymatic extension reactions, in which covalent incorporation of nucleotides to form the complement of the template is achieved by the action of DNA polymerase. Primers were attached via-C6H12SH,-C12H24SH, and -TTACAATC6H12SH linkers attached at the 5′ end. Primer coverage on the nanoparticles was varied by dilution with 5′HSC6H12AAA AAA3′. Hybridization efficiencies were determined as a function of linker length, primer coverage, complement length (12-mer vs 88-mer), and primer:complement concentration ratio. In all cases, hybridization for the 88-mer was less efficient than for the 12-mer. Low primer surface coverages, greater particle-primer separation, and higher primer:complement ratios led to optimal hybridization. Hybridization efficiencies as high as 98% and 75% were observed for the 12-mer and 88-mer, respectively. Enzymatic extension of particle-bound primers was observed under all conditions tested; however, the efficiency of the reaction was strongly affected by linker length and primer coverage. Extension of primers attached by the longest linker was as efficient as the solution-phase reaction.

Original languageEnglish (US)
Pages (from-to)7314-7323
Number of pages10
JournalJournal of the American Chemical Society
Volume124
Issue number25
DOIs
StatePublished - Jun 26 2002

All Science Journal Classification (ASJC) codes

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry

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