Cantilever-Based Optical Deflection Assay for Discrimination of DNA Single-Nucleotide Mismatches

Cantilever-Based Optical Deflection Assay for Discrimination of DNA Single-Nucleotide Mismatches. Hansen, K. M., Ji, H., Wu, G., Datar, R., Cote, R., Majumdar, A., & Thundat, T. Analytical Chemistry, 73:1567-1571, April 1, 2001, 2001.

Paper abstract bibtex

Characterization of single-nucleotide polymorphisms is a major focus of current genomics research. We demonstrate the discrimination of DNA mismatches using an elegantly simple microcantilever-based optical deflection assay, without the need for external labeling. Gold-coated silicon AFM cantilevers were functionalized with thiolated 20- or 25-mer probe DNA oligonucleotides and exposed to target oligonucleotides of varying sequence in static and flow conditions. Hybridization of 10-mer complementary target oligonucleotides resulted in net positive deflection, while hybridization with targets containing one or two internal mismatches resulted in net negative deflection. Mismatched targets produced a stable and measurable signal when only a four-base pair stretch was complementary to the probe sequence. This technique is readily adaptable to a high-throughput array format and provides a distinct positive/negative signal for easy interpretation of oligonucleotide hybridization.

@article {735,
	title = {Cantilever-Based Optical Deflection Assay for Discrimination of DNA Single-Nucleotide Mismatches},
	journal = {Analytical Chemistry},
	volume = {73},
	year = {2001},
	month = {April 1, 2001},
	pages = {1567-1571},
	abstract = {Characterization of single-nucleotide polymorphisms is a major focus of current genomics research. We demonstrate the discrimination of DNA mismatches using an elegantly simple microcantilever-based optical deflection assay, without the need for external labeling. Gold-coated silicon AFM cantilevers were functionalized with thiolated 20- or 25-mer probe DNA oligonucleotides and exposed to target oligonucleotides of varying sequence in static and flow conditions. Hybridization of 10-mer complementary target oligonucleotides resulted in net positive deflection, while hybridization with targets containing one or two internal mismatches resulted in net negative deflection. Mismatched targets produced a stable and measurable signal when only a four-base pair stretch was complementary to the probe sequence. This technique is readily adaptable to a high-throughput array format and provides a distinct positive/negative signal for easy interpretation of oligonucleotide hybridization.},
	isbn = {0003-2700},
	url = {http://dx.doi.org/10.1021/ac0012748},
	author = {Hansen, Karolyn M. and Ji, Hai-Feng and Wu, Guanghua and Datar, Ram and Cote, Richard and Majumdar, Arunava and Thundat, Thomas}
}

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