Gradient temperature hybridization using a thermocycler for RNase protection assays

The RNase protection assay (RPA) is a sensitive and quantitative assay for the detection of specific RNAs. Because some probes require an optimal temperature for efficient hybridization to the target RNAs, preliminary experiments to determine the optimal temperatures have to be done empirically, a process that is laborious and time-consuming. In order to simplify this process, a new gradient temperature hybridization procedure has been developed with the use of a thermocycler. This procedure eliminates the need for preliminary experiments to determine the optimal temperature of hybridization for a given probe.     

Integration of plasmonic heating and on‐chip temperature sensor for nucleic acid amplification assays

Nucleic acid tests have been widely used for diagnosis of diseases by detecting the relevant genetic markers that are usually amplified using polymerase chain reaction (PCR). This work reports the use of a plasmonic device as an efficient and low‐cost PCR thermocycler to facilitate nucleic acid‐based diagnosis. The thermoplasmonic device, consisting of a one‐dimensional metal grating, exploited the strong light absorption of plasmonic resonance modes to heat up PCR reagents using a near‐infrared laser source. The plasmonic device also integrated a thin‐film thermocouple on the metal grating to monitor the sample temperature. The plasmonic thermocycler is capable of performing a PCR amplification cycle in ~2.5 minutes. We successfully demonstrated the multiplex and real‐time PCR amplifications of the antibiotic resistance genes using the genomic DNAs extracted from Acinetobacter baumannii, Klebsiella pneumonia, Escherichia coli and Campylobacter.