RESUMO
Digital PCR (dPCR) is a technique for absolute quantification of nucleic acid molecules. To develop a dPCR technique that enables more accurate nucleic acid detection and quantification, we established a novel dPCR apparatus known as centrifugal force real-time dPCR (crdPCR). This system is efficient than other systems with only 2.14% liquid loss by dispensing samples using centrifugal force. Moreover, we applied a technique for analyzing the real-time graph of the each micro-wells and distinguishing true/false positives using artificial intelligence to mitigate the rain, a persistent issue with dPCR. The limits of detection and quantification were 1.38 and 4.19 copies/µL, respectively, showing a two-fold higher sensitivity than that of other comparable devices. With the integration of this new technology, crdPCR will significantly contribute to research on next-generation PCR targeting absolute micro-analysis.
Assuntos
DNA , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase em Tempo Real/métodos , DNA/análise , DNA/genética , Centrifugação/métodos , Limite de DetecçãoRESUMO
In this study, ultrathin and defect-free graphene (Gr) sheets were prepared through a fluid dynamics-induced shear exfoliation method using graphite. The high hydrophobicity and surface area of Gr make it attractive as a solid-contact ion-to-electron transducer for potentiometric K+ sensors, in which the electrodes are fabricated through a screen-printing process. The electrochemical characterization demonstrates that Gr solid contact results in a high double-layer capacitance, potential stability, and strong resistance against water layer, gases, and light. The Gr-based K+ sensors showed a Nernstian slope of 53.53â¯mV/log[K+] within a linear concentration range of 10-1-10-4 M, a low detection limit of 10-4.28â¯M, a fast response time of ~8â¯s, good repeatability, and excellent long-term stability. Moreover, the Gr-based K+ sensors provided accurate ion concentrations in actual samples of human sweat and sports drinks.