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Characterization of PNA and DNA Immobilization and Subsequent Hybridization with DNA Using Acoustic-Shear-Wave Attenuation Measurements

Fredrik Höök (Institutionen för teknisk fysik) ; Arghya Ray (Institutionen för fysikalisk kemi) ; Bengt Nordén (Institutionen för fysikalisk kemi) ; Bengt Kasemo (Institutionen för teknisk fysik)
Langmuir (0743-7463 ). Vol. 17 (2001), 26, p. 8305-8312.
[Artikel, refereegranskad vetenskaplig]

We report here how the quartz crystal microbalance with dissipation monitoring (QCM-D) technique, simultaneously measuring changes in the induced energy dissipation, D (cf. viscoelastic properties), and the frequency, f (cf. coupled mass), can be used to characterize the bound state of single-stranded peptide nucleic acid (PNA) and deoxyribose nucleic acid (DNA) in relation to their ability to function as selective probe(s) for fully complementary and single-mismatch DNA. The possibility to use the QCM-D technique for detection of binding kinetics and structural differences in the formed duplexes is also presented. We found that thiol-PNA and thiol-DNA attached via a sulfur group directly on a bare-gold surface are less efficient as probes for DNA than are biotin-PNA and biotin-DNA, coupled on top of a two-dimensional (2-D) arrangement of streptavidin, formed on a biotinylated phospholipid bilayer on a SiO2 surface. The fully complementary and singly mismatched DNA oligomers hybridize with the immobilized PNA and DNA. A single mismatch is discriminated via a significant difference in the binding and dissociation kinetics, demonstrating a high selectivity and thus successful immobilization of functional single strands. The observed ratios between hybridization-induced energy dissipation (DeltaD) and the frequency shift (Deltaf) made it possible to discriminate thiol-PNA directly attached to a gold surface from biotin-PNA coupled to the streptavidin 2-D arrangement, where the former were shown to be inefficient for detecting subsequent hybridization. Structural differences of the immobilized layers composed of biotin-PNA-DNA and biotin-DNA-DNA were clearly reflected by the DeltaD and Deltaf response.

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Denna post skapades 2011-01-05. Senast ändrad 2011-06-23.
CPL Pubid: 132471


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Institutioner (Chalmers)

Institutionen för teknisk fysik (1900-2015)
Institutionen för fysikalisk kemi (1900-2003)


Nanovetenskap och nanoteknik
Fysikalisk kemi

Chalmers infrastruktur