Selective colorimetric detection of polynucleotides based on the distance-dependent optical properties of gold nanoparticles

Science

Volumn 277, Issue 5329, 1997, Pages 1078-1081

  Elghanian, Robert ^a   Storhoff, James J ^a   Mucic, Robert C ^a   Letsinger, Robert L ^a   Mirkin, Chad A ^a  

^a NORTHWESTERN UNIVERSITY   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

OLIGONUCLEOTIDE;

ARTICLE; COLORIMETRY; COVALENT BOND; HYBRIDIZATION; NONHUMAN; OLIGONUCLEOTIDE PROBE; OPTICAL DENSITY; PRIORITY JOURNAL;

BIOSENSING TECHNIQUES; COLORIMETRY; GOLD; MICROCHEMISTRY; NUCLEIC ACID HYBRIDIZATION; OLIGONUCLEOTIDE PROBES; POLYDEOXYRIBONUCLEOTIDES; SPECTROPHOTOMETRY; TEMPERATURE;

EID: 0030768397     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.277.5329.1078     Document Type: Article

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17. Multiple loading of the oligonucleotides is necessary for subsequent cross-linking in our system. The surface chemistry involved in linking alkylthiols to Au surfaces is still a subject of debate [see C. S. Weisbecker et al., Langmuir 12, 3763 (1996)]. An alternative method of binding oligonucleotides covalently to Au particles, which was developed for a different purpose, was reported to give ∼1-nm nanoparticles containing a single oligomer on a particle [A. P. Alivisatos et al., Nature 382 609 (1996)].
18. Multiple loading of the oligonucleotides is necessary for subsequent cross-linking in our system. The surface chemistry involved in linking alkylthiols to Au surfaces is still a subject of debate [see C. S. Weisbecker et al., Langmuir 12, 3763 (1996)]. An alternative method of binding oligonucleotides covalently to Au particles, which was developed for a different purpose, was reported to give ∼1-nm nanoparticles containing a single oligomer on a particle [A. P. Alivisatos et al., Nature 382 609 (1996)].
19. Subsequent experiments have shown that the oligonucleotide spacer is not essential.
20. The concentrations for the three components were the same as those described in the caption for Fig. 3A.
21. For another example in which the rate of a reaction dependent on the hybridization of oligonucleotides was accelerated by the freezing of an aqueous solution of the components, see S. M. Gryaznov and R. L. Letsinger, J. Am. Chem. Soc. 115, 3808 (1993).
22. m for the nanoparticle system hybridized by the freeze-thaw method agreed (±0.2°C) with that for the nanoparticle system hybridized at room temperature (24 hours).
23. Transmission electron microscopy pictures of typical aggregates are available in supplementary material. Similar images can be found in (70).
24. For this experiment, 1 μl of solution containing 10 fmol of target oligonucleotide and 1 μl of solution containing both nanoparticle probes in a buffer (0.3
25. 18 TLC plate. A blue spot developed. In the absence of target, the color remained pink. For comparison, the lower limit for detecting an oligonucleotide with the use of a probe labeled with fluorescein in a sandwich hybridization system was reported to be 500 fmol [M. S. Urdea et al., Nucleic Acids Res. 16, 4937 (1988)].
26. We acknowledge support by grants from the National Institute of General Medical Sciences (GM 10265) and the Office of Naval Research (N0014-94-1-0703 and N00014-97-10430) and the Department of Defense (MURI DAAG 55-97-1-0133).