Assessment of Cost-Efficient Thermocycler Prototype for Polymerase Chain Reaction and Loop-Mediated Isothermal Amplification

DNA amplification-based diagnostic is the most accurate method among others, especially during the COVID-19 pandemic. Thus, increasing the global demand for instrumentation andamplification reagents locally, hence increasing import. It is a worrying state in terms of logisticsand the future domestic market. An effort for domestic production is a must. Previously a costefficientthermocycler prototype using Raspberry Pi and Phyton coding is constructed.Thermocycler prototype flow measurement and heat distribution have previously been tested. Thisresearch aims to compare thermocycler prototypes and commercial for in two types of DNAamplification reactions, polymerase chain reaction (PCR) and loop-mediated isothermalamplification (LAMP). PCR is the most and more common method than LAMP, with the maindifference of PCR require thermal cycling and LAMP operate in isothermal conditions. LAMP has aquicker reaction time and operates at a lower temperature. DNA pol with high strand displacementactivity is used for LAMP, in this research Bsm pol is used for LAMP and Taq pol for PCR. Since theprototype thermocycler is designed to be as simple and inexpensive as possible for ease ofmanufacture and accessibility for every layer of society. Hence, its heat control and stability are notas good as a commercial thermocycler, with huge temperature fluctuation resonance from its setpoint.That causes prototype incapability of performing PCR, no DNA band at 250-500 bp range ingel electrophoresis. However, the prototype is capable of performing LAMP, existing <100 bp DNAgradient band in gel electrophoresis. The prototype is also capable of performing LAMP below itsprotocol temperature and time separately, 62°C and 40 minutes compared to the protocol of 66°Cand 60 minutes