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.
Scuola Superiore di Catania, c/o Dipartimento di Scienze Chimiche, Università di Catania, Catania, Italy.
Laura Maria
LM
Zanoli
Marco
M
Licciardello
Roberta
R
D'Agata
Claudia
C
Lantano
Alessandro
A
Calabretta
Roberto
R
Corradini
Rosangela
R
Marchelli
Giuseppe
G
Spoto
D018744:DNA, Plant / Q000032:analysis / Q000235:genetics; D004277:DNA, Single-Stranded / Q000032:analysis / Q000235:genetics; D046210:Microfluidic Analytical Techniques / Q000379:methods*; D031658:Olea / Q000737:chemistry / Q000145:classification / Q000235:genetics*; D020135:Peptide Nucleic Acids / Q000737:chemistry* / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods*; D020641:Polymorphism, Single Nucleotide; D013025:Glycine max / Q000737:chemistry / Q000145:classification / Q000235:genetics*
D018744:DNA, Plant; D004277:DNA, Single-Stranded; D020135:Peptide Nucleic Acids