Strategies for optimizing DNA hybridization on surfaces

寡核苷酸 核酸 核酸热力学 DNA DNA–DNA杂交 杂交探针 分子信标 多核苷酸 化学 生物化学 基序列
作者
Hadi Ravan,Soheila Kashanian,Nima Sanadgol,Arastoo Badoei-Dalfard,Zahra Karami
出处
期刊:Analytical Biochemistry [Elsevier]
卷期号:444: 41-46 被引量:67
标识
DOI:10.1016/j.ab.2013.09.032
摘要

Specific and predictable hybridization of the polynucleotide sequences to their complementary counterparts plays a fundamental role in the rational design of new nucleic acid nanodevices. Generally, nucleic acid hybridization can be performed using two major strategies, namely hybridization of DNA or RNA targets to surface-tethered oligonucleotide probes (solid-phase hybridization) and hybridization of the target nucleic acids to randomly distributed probes in solution (solution-phase hybridization). Investigations into thermodynamic and kinetic parameters of these two strategies showed that hybridization on surfaces is less favorable than that of the same sequence in solution. Indeed, the efficiency of DNA hybridization on surfaces suffers from three constraints: (1) electrostatic repulsion between DNA strands on the surface, (2) steric hindrance between tethered DNA probes, and (3) nonspecific adsorption of the attached oligonucleotides to the solid surface. During recent years, several strategies have been developed to overcome the problems associated with DNA hybridization on surfaces. Optimizing the probe surface density, application of a linker between the solid surface and the DNA-recognizing sequence, optimizing the pH of DNA hybridization solutions, application of thiol reagents, and incorporation of a polyadenine block into the terminal end of the recognizing sequence are among the most important strategies for enhancing DNA hybridization on surfaces.
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