期刊
JOURNAL OF COLLOID AND INTERFACE SCIENCE
卷 630, 期 -, 页码 193-201出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.jcis.2022.10.018
关键词
Specific buffer effects; DNA; Melting temperature; Electrostatic interaction
This study evaluates the specific effects of buffers on the thermal stability of DNA, showing that increasing buffer concentration enhances DNA stability and different buffer types have different influences.
The specific effects of salts (strong electrolytes) on biomolecular properties have been investigated for more than a century. By contrast, the specific role of pH buffers (weak electrolytes and their salts) has usually been ignored. Here, specific buffer effects on DNA thermal stability were evaluated by measuring the melting curve of calf thymus DNA through UV-vis spectroscopy. The study was carried out using phosphate, Tris, citrate and cacodylate buffers at fixed pH 7.4 at concentrations varying systematically in the range 1-600 mM. DNA stability increases with buffer concentration and is influenced specifically by buffer type. To interpret empirical data, a theoretical model was applied with parameters quantifying the impact of buffer on the DNA backbone charge. Comparing the buffer effects via buffer ionic strength rather than buffer concentration, we find that the buffers stabilize DNA in the order Tris > cacodylate > phosphate > citrate.(c) 2022 Published by Elsevier Inc.
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