Peptide nucleic acid molecular beacons for the detection of PCR amplicons in droplet-based microfluidic devices(358 views) Zanoli LM, Licciardello M, D'Agata R, Lantano C, Calabretta A, Corradini R, Marchelli R, Spoto G
Scuola Superiore di Catania, C/o Dipartimento di Scienze Chimiche, Università di Catania, Viale Andrea Doria 6, Catania 95125, Italy
Dipartimento di Chimica Organica e Industriale, Università di Parma, V.le G. P. Usberti 17/a, Parma 43100, Italy
Istituto Biostrutture e Bioimmagini, CNR, Viale A. Doria 6, Catania 95125, Italy
Center for Molecular Medicine, Austrian Academy of Sciences, Lazarettgasse 14, Vienna 1090, Austria
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Peptide nucleic acid molecular beacons for the detection of PCR amplicons in droplet-based microfluidic devices
The use of droplet-based microfluidics and peptide nucleic acid molecular beacons for the detection of polymerase chain reaction (PCR)-amplified DNA sequences within nanoliter-sized droplets is described in this work. The nanomolar-attomolar detection capabilities of the method were preliminarily tested by targeting two different single-stranded DNA sequences from the genetically modified Roundup Ready soybean and the Olea europaea genomes and detecting the fluorescence generated by peptide nucleic acid molecular beacons with fluorescence microscopy. Furthermore, the detection of 10 nM solutions of PCR amplicon of DNA extracted from leaves of O. europaea L. encapsulated in nanoliter-sized droplets was performed to demonstrate that peptide nucleic acid molecular beacons can discriminate O. europaea L. cultivar species carrying different single-nucleotide polymorphisms.
Peptide nucleic acid molecular beacons for the detection of PCR amplicons in droplet-based microfluidic devices