A DNA analysis device based on poly(methyl methacrylate) (PMMA) microfluidic chips was developed. A PMMA chip with cross microchannels was fabricated by a simple hot embossing. Microchannels were modified with a static adsorptive coating methodusing 2% hydroxyethyl cellulose. A high-voltage power unit, variable in the range 0-1800 V, was used for onchip DNA sample injection and gel electrophoretic separation. High speed, high resolution DNA analysis was obtained with the home-built PMMA chip in asieving matrix containing 2% hydroxy-ethyl cellulose with a blue intercalating dye, TO-PRO-3(TP3), by using diode laser induced fluorescence detection based on optical fibers with a 670 nm long-pass filter. The DNA analysis device was applied for the separation of X-174/Hae Ⅲ DNA digest sample with 11 fragments ranging from 72 to 1353 bp. A separation efficiency of 1.14×10⁶ plates/m was obtained for the 603 bp fragments, while the R of 271/281 bp fragments was 1.2. The device was characterized by simple design, low cost for fabrication and operation, reusable PMMA chips,and good reproducibility. A portable microfluidic device for DNA analysis can be developed for clinical diagnosis and disease screening.