We proposed a novel method of fabricating polydimethylsiloxane (PDMS) microfluidic chip polymer master molds in this paper. The method mainly includes two steps. First, a stainless steel slice was laser etched to form a metal model. Then, the organic solution of poly(methyl methacrylate) (PMMA) was casted onto the metal model to fabricate the PMMA master which subsequently would be used to fabricate PDMS chips. We systematically researched different laser parameters influencing the surface status of microchannels and obtained optimized etching parameters. We investigated and optimized the organic solution composition of PMMA while casting chip masters, and developed a method to form fine polymer masters using two different viscosity solutions to cast the model in turn, and studied the repeatable replication. Then, we investigated physical performance of this chip and evaluated the practicability by analyzing Rhodamine B. Compared with present methods, the proposed method does not need photolithography on photoresistant and chemical etching. The entire fabricating progress is simple, fast, low-cost and can be controlled easily. Only several minutes are required to make a metal model, 3 hours for a PMMA master, and one day for PDMS chips. We propose a novel method of fabricating polydimethylsiloxane (PDMS) microfluidic chip polymer master molds in this paper. The method mainly includes two steps. First, a stainless steel slice was laser etched to form a metal model. Then, the organic solution of poly methyl methacrylate (PMMA) was casted onto the metal model to fabricate the PMMA master which subsequently would be used to fabricate PDMS chips. We systematically researched different laser parameters influencing the surface status of microchannels and obtained optimized etching parameters. We investigated and optimized the organic solution composition of PMMA while casting chip masters, and developed a method to form fine polymer masters using two different viscosity solutions to cast the model in turn, and studied the repeatable replication. Then, we investigated physical performance of this chip and evaluated the practicability by analyzing Rhodamine B. Compared with present methods, the proposed method does not need The entire fabricating progress is simple, fast, low-cost and can be controlled easily. Only several minutes are required to make a metal model, 3 hours for a PMMA master, and one day for PDMS chips.