Poly(dimethylsiloxane) (PDMS) microstructures have been widely used in bio-microelectromechanical systems (bio-MEMS) for various types of analytical, diagnostic and therapeutic applications. However, PDMS-based soft lithographic techniques still use conventional microfabrication processes to generate a master mold, which requires access to clean room facilities and costly equipment. With the increasing use of these systems in various fields, the development of benchtop systems for fabricating microdevices is emerging as an important challenge in their widespread use. Here we demonstrate a simple, low-cost and rapid method to fabricate PDMS microstructures by using micropatterned poly(ethylene glycol) diacrylate (PEGDA) master molds. In this method, PEGDA microstructures were patterned on a glass substrate by photolithography under ambient conditions and by using simple tools. The resulting PEGDA structures were subsequently used to generate PDMS microstructures by standard molding in a reproducible and repeatable manner. The thickness of the PEGDA microstructures was controllable from 15 to 300 µm by using commonly available spacer materials. We also demonstrate the use of this method to fabricate microfluidic channels capable of generating concentration gradients. In addition, we fabricated PEGDA microstructures by photolithography from the light generated from commonly available laminar cell culture hood. These data suggest that this approach could be beneficial for fabricating low-cost PDMS-based microdevices in resource limited settings.