An advanced analytical chemistry undergraduate laboratory module on microfluidics that spans 4 weeks (4 h per week) is presented. The laboratory module focuses on comprehensive experiential learning of microfluidic device fabrication and the core characteristics of microfluidic devices as they pertain to fluid flow and the manipulation of samples. Experiments include the cleanroom-free fabrication of glass-polydimethylsiloxane (PDMS) microfluidic devices by use of thermoplastic molds and soft lithography, determination of the zeta potential at the microchannel walls, investigations of electroosmotic flow, determination of Péclet numbers, and diffusion coefficients based on diffusion-limited reagent mixing. The module concludes with the development of a microchannel electrophoresis lab-on-a-chip device with in-line fluorimetric detection for the separation of a DNA ladder. This laboratory module has been offered for two consecutive years and has received favorable student feedback.