Characterization of DNA immobilization and subsequent hybridization using in situ quartz crystal microbalance, fluorescence spectroscopy, and surface plasmon resonance

Yoon Kyoung Cho; Sunhee Kim; Young A. Kim; Hee Kyun Lim; Kyusang Lee; Daesung Yoon; Geunbae Lim; Y. Eugene Pak; Tai Hwan Ha; Kwan Kim

doi:10.1016/j.jcis.2004.05.008

Characterization of DNA immobilization and subsequent hybridization using in situ quartz crystal microbalance, fluorescence spectroscopy, and surface plasmon resonance

Yoon Kyoung Cho, Sunhee Kim, Young A. Kim, Hee Kyun Lim, Kyusang Lee, Daesung Yoon, Geunbae Lim, Y. Eugene Pak, Tai Hwan Ha, Kwan Kim

Research output: Contribution to journal › Article › peer-review

49 Citations (Scopus)

Abstract

We have characterized the immobilization of thiol-modified oligomers on Au surfaces and subsequent hybridization with a perfectly matched or single-base mismatched target using a quartz crystal microbalance (QCM) and fluorescence spectroscopy. The surface density of immobilized probe molecules and the hybridization efficiency depending on the type of buffer and salt concentration were investigated. We observed some ambiguities in surface coverage deduced from QCM measurement and adopted a complementary fluorescence displacement method. Direct comparison of surface coverage deduced from frequency change in QCM measurement and determined by the fluorescence exchange reaction revealed that QCM results are highly overestimated and the amount of overestimation strongly depends on the type of buffer and the structure of the film. Discrimination capability of the surface attached 15-mer probe was also examined using a single-base mismatched target at various hybridization temperatures. Hybridization efficiency depending on the type of single base mismatch was investigated using surface plasmon resonance (SPR).

Original language	English
Pages (from-to)	44-52
Number of pages	9
Journal	Journal of Colloid and Interface Science
Volume	278
Issue number	1
DOIs	https://doi.org/10.1016/j.jcis.2004.05.008
Publication status	Published - 2004 Oct 1
Externally published	Yes

Bibliographical note

Funding Information:
We thank Dongkyu Jin for providing us the sequence of DNA samples and Taejoon Kwon for calculating the melting temperature of oligonucleotides using the nearest-neighbor model. This work was supported by the Ministry of Commerce, Industry and Energy (MOCIE) of the Republic of Korea under the a next generation new technology development project (00008069) through the Digital Bio Laboratory at the Samsung Advanced Institute of Technology (SAIT).

Copyright:
Copyright 2008 Elsevier B.V., All rights reserved.

ASJC Scopus subject areas

Electronic, Optical and Magnetic Materials
Biomaterials
Surfaces, Coatings and Films
Colloid and Surface Chemistry

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10.1016/j.jcis.2004.05.008

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Cho, Y. K., Kim, S., Kim, Y. A., Lim, H. K., Lee, K., Yoon, D., Lim, G., Pak, Y. E., Ha, T. H., & Kim, K. (2004). Characterization of DNA immobilization and subsequent hybridization using in situ quartz crystal microbalance, fluorescence spectroscopy, and surface plasmon resonance. Journal of Colloid and Interface Science, 278(1), 44-52. https://doi.org/10.1016/j.jcis.2004.05.008

Characterization of DNA immobilization and subsequent hybridization using in situ quartz crystal microbalance, fluorescence spectroscopy, and surface plasmon resonance. / Cho, Yoon Kyoung; Kim, Sunhee; Kim, Young A. et al.
In: Journal of Colloid and Interface Science, Vol. 278, No. 1, 01.10.2004, p. 44-52.

Research output: Contribution to journal › Article › peer-review

Cho, YK, Kim, S, Kim, YA, Lim, HK, Lee, K, Yoon, D, Lim, G, Pak, YE, Ha, TH & Kim, K 2004, 'Characterization of DNA immobilization and subsequent hybridization using in situ quartz crystal microbalance, fluorescence spectroscopy, and surface plasmon resonance', Journal of Colloid and Interface Science, vol. 278, no. 1, pp. 44-52. https://doi.org/10.1016/j.jcis.2004.05.008

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abstract = "We have characterized the immobilization of thiol-modified oligomers on Au surfaces and subsequent hybridization with a perfectly matched or single-base mismatched target using a quartz crystal microbalance (QCM) and fluorescence spectroscopy. The surface density of immobilized probe molecules and the hybridization efficiency depending on the type of buffer and salt concentration were investigated. We observed some ambiguities in surface coverage deduced from QCM measurement and adopted a complementary fluorescence displacement method. Direct comparison of surface coverage deduced from frequency change in QCM measurement and determined by the fluorescence exchange reaction revealed that QCM results are highly overestimated and the amount of overestimation strongly depends on the type of buffer and the structure of the film. Discrimination capability of the surface attached 15-mer probe was also examined using a single-base mismatched target at various hybridization temperatures. Hybridization efficiency depending on the type of single base mismatch was investigated using surface plasmon resonance (SPR).",

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N2 - We have characterized the immobilization of thiol-modified oligomers on Au surfaces and subsequent hybridization with a perfectly matched or single-base mismatched target using a quartz crystal microbalance (QCM) and fluorescence spectroscopy. The surface density of immobilized probe molecules and the hybridization efficiency depending on the type of buffer and salt concentration were investigated. We observed some ambiguities in surface coverage deduced from QCM measurement and adopted a complementary fluorescence displacement method. Direct comparison of surface coverage deduced from frequency change in QCM measurement and determined by the fluorescence exchange reaction revealed that QCM results are highly overestimated and the amount of overestimation strongly depends on the type of buffer and the structure of the film. Discrimination capability of the surface attached 15-mer probe was also examined using a single-base mismatched target at various hybridization temperatures. Hybridization efficiency depending on the type of single base mismatch was investigated using surface plasmon resonance (SPR).

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Characterization of DNA immobilization and subsequent hybridization using in situ quartz crystal microbalance, fluorescence spectroscopy, and surface plasmon resonance

Abstract

Bibliographical note

ASJC Scopus subject areas

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