Rapid and quantitative detection of the binding of nucleic acids to surface-immobilized probes remains a challenge in many biomedical applications. We investigated the hybridization of a set of fully complementary and defected 12-base long DNA oligomers by using the Reflective Phantom Interface (RPI), a recently developed multiplexed label-free detection technique. Based on the simple measurement of reflected light intensity, this technology enables to quantify the hybridization directly as it occurs on the surface with a sensitivity of 10 pg mm-2. We found a strong effect of single-base mismatches and of their location on hybridization kinetics and equilibrium binding. In line with previous studies, we found that DNA-DNA binding is weaker on a surface than in the bulk. Our data indicate that this effect is a consequence of weak nonspecific binding of the probes to the surface.

Label-free detection of DNA single-base mismatches using a simple reflectance-based optical technique / G. Nava, E. Ceccarello, F. Giavazzi, M. Salina, F. Damin, M. Chiari, M. Buscaglia, T. Bellini, G. Zanchetta. - In: PHYSICAL CHEMISTRY CHEMICAL PHYSICS. - ISSN 1463-9076. - 18:19(2016), pp. 13395-13402.

Label-free detection of DNA single-base mismatches using a simple reflectance-based optical technique

G. Nava
Primo
;
E. Ceccarello
Secondo
;
F. Giavazzi;M. Salina;M. Buscaglia;T. Bellini
Penultimo
;
G. Zanchetta
2016

Abstract

Rapid and quantitative detection of the binding of nucleic acids to surface-immobilized probes remains a challenge in many biomedical applications. We investigated the hybridization of a set of fully complementary and defected 12-base long DNA oligomers by using the Reflective Phantom Interface (RPI), a recently developed multiplexed label-free detection technique. Based on the simple measurement of reflected light intensity, this technology enables to quantify the hybridization directly as it occurs on the surface with a sensitivity of 10 pg mm-2. We found a strong effect of single-base mismatches and of their location on hybridization kinetics and equilibrium binding. In line with previous studies, we found that DNA-DNA binding is weaker on a surface than in the bulk. Our data indicate that this effect is a consequence of weak nonspecific binding of the probes to the surface.
biosensors; DNA; label-free; optical sensors
Settore FIS/07 - Fisica Applicata(Beni Culturali, Ambientali, Biol.e Medicin)
   Next Generation Biomimetic Analytical Platforms. Building Blocks for Ubiquitous Environmental Sensor Networks
   NAPES
   EUROPEAN COMMISSION
   FP7
   604241
2016
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/421559
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