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Early diagnosis of pathogens is key to reducing their spreading and to preventing severe health risks. The COVID-19 pandemic showed us the need for rapid point-of-care tests. Here, we describe the preparation of an amplification master mix for point-of-care diagnostics. Therefore, two off-patent amplification enzymes were designed, expressed and purified. The preparation of the key components enables independence from delivery issues, manufacturer portfolio and product information's. For long-term storage and cold-free transport, our fabricated amplification mix was lyophilised. Finally, we applied our lyophilised master mix on an integrated point-of-care diagnostic system and could detect 10 copies/μL COVID-19 RNA. The combination of stable, cold-free reagents with the mobile and low-cost device will allow molecular diagnostics of pathogens in a field or home setting.