Layer-by-layer assemblies based on deoxyribonucleic acid (DNA) hybridization have potential for various bio- and nanotechnology applications because of their programmability, biodegradability, and ability to control the structure of the assemblies on the nanometer scale. Herein, we investigate the growth and salt stability of DNA films by the optical technique dual polarization interferometry and numerically model the film buildup. The DNA films were assembled by sequentially depositing pairs of oligonucleotides comprised of two different block sequences onto the surface. The oligonucleotides used in the assembly of the different films were as follows: a homopolymeric diblock pair of AxGx/TxC x (x = 15, 20, or 30), a homopolymeric diblock pair of A 15G15/C15T15 in which the orientation of the T15C15 diblock was reversed (C 15T15), and a random diblock pair of X15Y 15/X'15Y'15. The characteristics of the layer growth were highly dependent on the type of the oligonucleotide pair used: the mass of DNA deposited followed a linear, stepwise, or saturated growth with increasing layer deposition. The layer growth of each film was numerically modeled by taking into account the effective hybridization rate and the effective dissociation rate of the oligonucleotides. The proposed modeling offers a framework for molecularly designing oligonucleotide pairs to obtain DNA multilayer films with desired physicochemical properties (thickness, density, stability).