Polydimethylsiloxane (PDMS) is a long‐standing material of significant interest in microfluidics due to its unique features. As such, rapid prototyping of PDMS‐based microchannels is of great interest. The most prevalent and conventional method for fabrication of PDMS‐based microchips relies on softlithography, the main drawback of which is the preparation of a master mold, which is costly and time‐consuming. To prevent the attachment of PDMS to the master mold, silanization is necessary, which can be detrimental for cellular studies. Additionally, using coating the mold with a cell‐compatible surfactant adds extra preprocessing time. Recent advances in 3D printing have shown great promise in expediting microfabrication. Nevertheless, current 3D printing techniques are sub‐optimal for PDMS softlithography. The feasibility of producing master molds suitable for rapid softlithography is demonstrated using a newly developed 3D‐printing resin. Moreover, the utility of this technique is showcased for a number of widely used applications, such as concentration gradient generation, particle separation, cell culture (to show biocompatibility of the process), and fluid mixing. This can open new opportunities for biologists and scientists with minimum knowledge of microfabrication to build functional microfluidic devices for their basic and applied research.