High-precision micromilling was employed as a cost-efficient method preparation of metal masters useful in fabrication of polymer microfluidic devices through replication techniques. In first application, a brass mold master was used for hot embossing of microchip electrophoresis devices in poly(methyl methacrylate) (PMMA). The sidewalls of the milled microstructures were characterized by a maximum average roughness (Ra) of 110 nm and mean peak height (Rpm) of 320 nm. SEM imaging showed a transfer of the sidewall roughness from the molding tool to the polymer microdevice. The electroosmotic flow (EOF) values for micromilled-based microchannels were comparable to ones in the LiGA-prepared devices (sidewall Ra = 20 nm) with values of ca. 3.7 x 10-4 cm2V-1s-1 (20 mM TBE buffer, pH 8.2), indicating insignificant effects of wall roughness on the bulk EOF. Numerical simulations showed that the additional volumes present in an injection cross due to curvature of the corners produced by micromilling lead to elongated sample plugs. PMMA microchip electrophoresis devices were used for a separation of pUC19 Sau3AI double-stranded DNA. The plate numbers achieved exceeded 1 million m-1 and were comparable to the plate numbers for the LiGA-based devices of similar geometry. In second application brass master was used as tool for preparation of poly(dimethylsiloxane) PDMS stencils for patterning of DNA microarrays onto a PMMA substrate. Four zip code probes immobilized onto the PMMA surface directed allele-specic ligation products containing mutations in the KRAS2 gene (12.2D, 12.2A, 12.2V, and 13.4D) to the appropriate address of a universal array with minimal amounts of crosshybridization or misligation.