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

Fingerprint

oligonucleotides
Oligonucleotides
Monolayers
polymerase chain reaction
Polymerase chain reaction
Molecules
probes
reaction products
disulfides
molecules
immobilization
Sulfhydryl Compounds
thiols
Gold
Disulfides
cancer
gold
Single-Stranded DNA
Surface plasmon resonance
Reaction products

Keywords

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

Cite this

Vikholm-Lundin, Inger ; Auer, Sanna ; Munter, Tony ; Fiegl, Heidi ; Apostolidou, Sophia. / Hybridization of binary monolayers of single-stranded oligonucleotides and short blocking molecules. In: Surface Science. 2009 ; Vol. 603, No. 4. pp. 620-624.
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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.",
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Vikholm-Lundin, I, Auer, S, Munter, T, Fiegl, H & Apostolidou, S 2009, 'Hybridization of binary monolayers of single-stranded oligonucleotides and short blocking molecules', Surface Science, vol. 603, no. 4, pp. 620-624. https://doi.org/10.1016/j.susc.2008.12.026

Hybridization of binary monolayers of single-stranded oligonucleotides and short blocking molecules. / Vikholm-Lundin, Inger (Corresponding Author); Auer, Sanna; Munter, Tony; Fiegl, Heidi; Apostolidou, Sophia.

In: Surface Science, Vol. 603, No. 4, 2009, p. 620-624.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Hybridization of binary monolayers of single-stranded oligonucleotides and short blocking molecules

AU - Vikholm-Lundin, Inger

AU - Auer, Sanna

AU - Munter, Tony

AU - Fiegl, Heidi

AU - Apostolidou, Sophia

PY - 2009

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N2 - 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.

AB - 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.

KW - oligonucleotides

KW - immobilization

KW - self-assembly

KW - hybridization

KW - surface plasmon resonance

U2 - 10.1016/j.susc.2008.12.026

DO - 10.1016/j.susc.2008.12.026

M3 - Article

VL - 603

SP - 620

EP - 624

JO - Surface Science

JF - Surface Science

SN - 0039-6028

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ER -