This paper reports a numerical and experimental study of acoustic streaming and micromixing in polydimethylsiloxane microchannels. The mixing between two fluids flowing in microchannels was evaluated through the following conditions: (1) using a 28 μm thick β-poly(vinylidene fluoride) (β-PVDF) as a piezoelectric transducer actuated with a 24 Vpp and 40 MHz sinusoidal voltage; (2) using different flow rates. The results suggest that the mixing length increases as the flow rate increases and that the acoustic streaming phenomenon leads to a reduction on the mixing length. The good qualitative agreement between numerical and experimental results is a valuable indicator to predict the mixing performance of microfluidic devices, for improving biological fluid analysis in diagnosis lab-on-a-chip devices.