The rs320 polymorphism of lipoprotein lipase gene has been reported to be associated with triglycerides, total cholesterol, HDL-C, and LDL-C. The study aimed to optimize the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method with primers, thermal cycles, and restriction enzyme which were selected and tested to identify opimal condition of PCR-RFLP method.