Hybridization of binary monolayers of single-stranded oligonucleotides and short blocking molecules

Inger Vikholm-Lundin (Corresponding Author), Sanna Auer, Tony Munter, Heidi Fiegl, Sophia Apostolidou

Research output: Contribution to journalArticleScientificpeer-review

8 Citations (Scopus)

Abstract

We have studied the immobilization of single stranded (ss) DNA oligonucleotides of 16–27 base pairs on gold. The oligonucleotides were thiol-modified (SH-ssDNA) or disulfide-modified via a dimethoxytrityl-group (DMT-S-S-ssDNA). Immobilization was performed by adsorption of the probes on the gold surface for 10–15 min, a time within which saturation coverage was obtained for both thiol- and disulfide-modified probes. Hereafter the layer was post-treated with hydroxyalkyl substituted lipoamides also for a time of 10–15 min. The surface density of layers with shorter probes (16–18 mer) was twice (2.4 ± 0.2 × 1013 probes/cm2) that of the longer probes (25–27 mer) as studied with surface plasmon resonance. Hybridization of single stranded polymerase chain reaction (PCR) amplified products with a length above 300 base pairs gave a very low hybridization response. For amplicons with about 100 base pairs the response was high. The surface coverage was comparable to that of complementary ssDNA binding (3.0 × 1012 strands/cm2). Surfaces made from SH-ssDNA showed a 30% higher hybridization response than surfaces made from DMT-S-S-ssDNA. The PCR amplified products used are of relevance in breast cancer diagnosis. The results clearly demonstrate that the single stranded PCR products might be used in label-free cancer diagnostics.
Original languageEnglish
Pages (from-to)620-624
JournalSurface Science
Volume603
Issue number4
DOIs
Publication statusPublished - 2009
MoE publication typeA1 Journal article-refereed

Keywords

  • oligonucleotides
  • immobilization
  • self-assembly
  • hybridization
  • surface plasmon resonance

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