The initial multi-mode interfacial velocity and density perturbations present at the onset of a small-Atwood-number, incompressible, miscible Rayleigh–Taylor instability- driven mixing layer have been quantified using a combination of experimental techniques. The streamwise interfacial and spanwise interfacial perturbations were measured using high-resolution thermocouples and planar laser-induced fluorescence (PLIF), respectively. The initial multi-mode streamwise velocity perturbations at the two-fluid density interface were measured using particle-image velocimetry (PIV). It was found that the measured initial conditions describe an initially anisotropic state, in which the perturbations in the streamwise and spanwise directions are independent of one another. The evolution of various fluctuating velocity and density statistics, together with velocity and density variance spectra, were measured using PIV and high-resolution thermocouple data. The evolution of the velocity and density statistics is used to investigate the early-time evolution and the onset of strongly nonlinear, transitional dynamics within the mixing layer. The early-time evolution of the density and vertical velocity variance spectra indicate that velocity fluctuations are the dominant mechanism driving the instability development. The implications of the present experimental measurements on the initialization of Reynolds-averaged turbulent transport and mixing models and of direct and large-eddy simulations of Rayleigh–Taylor instability-induced turbulence are discussed.