Native characterization of nucleic acid motif thermodynamics via non-covalent catalysis

作者：Wang Chunyan; Bae Jin H; Zhang David Yu

来源：Nature Communications, 2016, 7(1): 10319.

DOI：10.1038/ncomms10319

DNA hybridization thermodynamics is critical for accurate design of oligonucleotides for biotechnology and nanotechnology applications, but parameters currently in use are inaccurately extrapolated based on limited quantitative understanding of thermal behaviours. Here, we present a method to measure the Delta G degrees of DNA motifs at temperatures and buffer conditions of interest, with significantly better accuracy (6- to 14-fold lower s.e.) than prior methods. The equilibrium constant of a reaction with thermodynamics closely approximating that of a desired motif is numerically calculated from directly observed reactant and product equilibrium concentrations; a DNA catalyst is designed to accelerate equilibration. We measured the Delta G degrees of terminal fluorophores, single-nucleotide dangles and multinucleotide dangles, in temperatures ranging from 10 to 45 degrees C.

• 出版日期2016-1