Kinetics Study of DNA Hybridization by Using Atomic Force Microscopy and Surface Plasmon Resonance
Date Issued
2007
Date
2007
Author(s)
Huang, Po-Hsun
DOI
en-US
Abstract
Attempting to probe the specific intermolecular forces is a challenging task in molecular and structural biology. However, it is of great interest because such forces dominate the behavior of all materials and biological systems. We used atomic force microscopy (AFM) and surface plasmaon resonance (SPR) to study the specific interaction between the linear DNA and their target DNA (complementary and one-base mutation) in this work. At the same time, we also used the hybridization buffer solutions in two different concentrations to realize whether the ionic strength affects the hybridization between the single-stranded DNA. The Poisson statistical method was used to analyze the rupture force of individual molecule, the mean number of bonds between linear DNA and their target DNA, and the possible non-specific and long-range interactions. The average rupture force for single-base mismatch and complementary system were 0.20833 ± 0.07317 nN and 0.26296 ± 0.07638 nN, respectively. The individual rupture force of double-stranded DNA for the complementary system was also determined to be 0.07843 nN. Finally, we carried out the study of the binding kinetics between linear DNA and their target DNA by SPR. According to our experimental results, AFM and SPR succeeded in distinguishing the difference between totally complementary double-stranded DNA and complement with single-pair mismatch. Besides, it was verified that the hybridization buffer with high ionic strength would be helpful to the DNA hybridization.
Subjects
去氧核糖核酸
掃描探針顯微術
原子力顯微術
表面電漿共振
雜交
全反射
自組裝單分子膜
DNA
Scanning Probe Microscopy
Atomic Force Microscopy
Surface Plasmon Resonance
Hybridization
Total Internal Reflection
Self-Assembled Monolayers
Type
thesis
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