Chapter title |
Single-molecule optical-trapping measurements with DNA anchored to an array of gold nanoposts.
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Chapter number | 18 |
Book title |
Spectroscopic Methods of Analysis
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Published in |
Methods in molecular biology, January 2012
|
DOI | 10.1007/978-1-61779-806-1_18 |
Pubmed ID | |
Book ISBNs |
978-1-61779-805-4, 978-1-61779-806-1
|
Authors |
Paik DH, Perkins TT, D. Hern Paik, Thomas T. Perkins, Paik, D. Hern, Perkins, Thomas T. |
Abstract |
Gold-thiol chemistry is one of the most successful chemistries for conjugating biomolecules to surfaces, but such chemistry has not been exploited in optical-trapping experiments because of laser-induced ablation of gold. In this work, we describe a method to combine these two separate technologies without undue heating using DNA anchored to gold nanostructures (r = 50-250 nm; h ≈ 20 nm). Moreover, we demonstrate a quantitative and mechanically robust (>100 pN) optical-trapping assay. By using three dithiol phosphoramidites (DTPAs) incorporated into a polymerase chain reaction (PCR) primer, the gold-DNA bond remained stable in the presence of excess thiolated compounds. This chemical robustness allowed us to reduce nonspecific sticking by passivating the unreacted gold with methoxy-(polyethylene glycol)-thiol (mPEG-SH). Overall, this surface conjugation of biomolecules onto an ordered array of gold nanostructures by chemically and mechanically robust bonds provides a unique way to carry out spatially controlled, repeatable measurements of single molecules. |
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