The invention belongs to the field of
biomedicine, and discloses a
nucleic acid phase diagram analysis method based on a variable-temperature
ultraviolet absorption spectrum, which comprises the following steps: preparing buffer solutions with different gradient pH-
ionic strength combinations, mixing the buffer solutions with a target structure, performing an annealing and melting procedure, and continuously collecting
ultraviolet absorption spectrum data; matching the data of the temperature and the
absorbance changing along with the time according to the time to obtain the data of the
absorbance changing along with the temperature under the full
wavelength; fitting and normalizing
absorbance at the characteristic peak to obtain temperature dependence forming coefficients at different pH values and
ion strengths, grouping the temperature dependence forming coefficients, and performing transposition treatment to obtain an
ion strength dependence forming coefficient and a
pH dependence forming coefficient; a
single variable is fixed, and a temperature correlation
phase diagram, a pH correlation
phase diagram and an
ion strength correlation phase diagram are drawn by utilizing dependency forming coefficients of other two variables, so that the limitation that the existing
analysis method cannot reflect the stability of a
nucleic acid structure in a complex environment is overcome.