The current study introduces novel methodologies in forensic and diagnostic science, notably the isothermal amplification of STR loci via Recombinase Polymerase Amplification (RPA) and the development of RPA-compatible, probe-based genotyping. At the heart of this research is the isothermal mechanism of RPA, which facilitates STR amplification at a constant temperature, thus eliminating the need for the thermal cycling required in traditional Polymerase Chain Reaction (PCR) methods. Moreover, this study replaces the conventional requirement for allele detection, typically achieved through electrophoresis or massively parallel sequencing, with a lab-on-a-chip compatible, probe-based quantitative RPA (qRPA) detection method. This advancement reduces the need for extensive equipment, enhancing the feasibility of implementing this technology in field-deployable, disposable microfluidic lab-on-a-chip systems. Such an innovation holds significant potential for transformative applications, particularly in situations demanding rapid decision-making. Its utility extends to a range of fields, including rapid diagnostics, urgent crime scene investigations, immediate disaster victim identification, and critical food safety assessments.