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2014 ; 8
(5
): 4488-99
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Mechanisms of surface-mediated DNA hybridization
#MMPMID24708278
Monserud JH
; Schwartz DK
ACS Nano
2014[May]; 8
(5
): 4488-99
PMID24708278
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Single-molecule total internal reflection fluorescence microscopy was employed in
conjunction with resonance energy transfer (RET) to observe the dynamic behavior
of donor-labeled ssDNA at the interface between aqueous solution and a solid
surface decorated with complementary acceptor-labeled ssDNA. At least 100,000
molecular trajectories were determined for both complementary strands and
negative control ssDNA. RET was used to identify trajectory segments
corresponding to the hybridized state. The vast majority of molecules from
solution adsorbed nonspecifically to the surface, where a brief two-dimensional
search was performed with a 7% chance of hybridization. Successful hybridization
events occurred with a characteristic search time of ?0.1 s, and unsuccessful
searches resulted in desorption from the surface, ultimately repeating the
adsorption and search process. Hybridization was reversible, and two distinct
modes of melting (i.e., dehybridization) were observed, corresponding to
long-lived (?15 s) and short-lived (?1.4 s) hybridized time intervals. A strand
that melted back onto the surface could rehybridize after a brief search or
desorb from the interface. These mechanistic observations provide guidance for
technologies that involve DNA interactions in the near-surface region, suggesting
a need to design surfaces that both enhance the complex multidimensional search
process and stabilize the hybridized state.
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