Homogenous measurement during a circulation-water-based ultrahigh-speed polymerase chain reaction and melting curve analysis device

Abstract

Polymerase chain reaction (PCR) is an essential technique for almost all life science fields that amplifies few target DNA copies. We developed a rapid real-time microdroplet PCR device by rapid switching of two types of circulation hot water, a µL-sized droplet, and a thin film aluminum chip. Using this device, rapid PCR amplification was observed successfully and accomplished within 3 min. By real-time PCR, many samples could be measured simultaneously. Moreover, melting curve analysis is essential for detecting differences of PCR amplicons in detail such as those related to nonspecific amplification, single nucleotide polymorphism, and genotyping methods. Thus, we developed a device that can be used for melting curve analysis using circulation water. From the obtained results, temperature homogeneity on the reaction plate was maintained during PCR and melting curve analysis using high-speed circulation water. The results indicate that this system can be applied to various life science fields.

Navigating the Pandemic Response Life Cycle: Molecular Diagnostics and Immunoassays in the Context of COVID-19 Management

“…Fast DNA polymerases, engineered with higher processing rates, extension rates, and faster activation times were used with the rapid thermal cyclers [88] . Efforts to decrease PCR times to less than 5 min were achieved by instrumentation [89] , including systems using hot and cool water for heat transfer through metal block or thin plates [90] [70] [91] and infrared-mediated temperature cycling, which enabled 3 sec PCR cycles [92]. When glass capillaries were submerged in liquid gallium, PCR cycle time was further improved down to 2.7 sec [65].…”