Dyes as bifunctional markers of DNA hybridization on surfaces and mutation detection.
The interaction of small molecules with DNA has found diagnostic and therapeutic applications. In this work, we propose the use of two different dyes, in particular Azure A and Safranine, as bifunctional markers of on-surface DNA hybridization and potent tools for screening of specific gene mutations directly in real DNA PCR amplicons extracted from blood cells. By combining spectroscopic and electrochemical methods we demonstrate that both dyes can interact with single and double stranded DNA to a different extent, allowing reliable hybridization detection. From these data, we have also elucidated the nature of the interaction. We conclude that the binding mode is fundamentally intercalative with an electrostatic component. The dye fluorescence allows their use as nucleic acid stains for the detection of on-surfaces DNA hybridization. Its redox activity is exploited in the development of selective electrochemical DNA biosensors.
Departamento Química Analítica y Análisis Instrumental, Universidad Autónoma de Madrid, 28049, Madrid, Spain; Instituto Madrileño de Estudios Avanzados (IMDEA) Nanociencia, Ciudad Universitaria de Cantoblanco, 28049, Madrid, Spain.
Tania
T
García-Mendiola
Departamento Química Analítica y Análisis Instrumental, Universidad Autónoma de Madrid, 28049, Madrid, Spain.
María Ramos
MR
Cerro
Departamento Química Analítica y Análisis Instrumental, Universidad Autónoma de Madrid, 28049, Madrid, Spain.
José María
JM
López-Moreno
Departamento Química Analítica y Análisis Instrumental, Universidad Autónoma de Madrid, 28049, Madrid, Spain; Instituto Madrileño de Estudios Avanzados (IMDEA) Nanociencia, Ciudad Universitaria de Cantoblanco, 28049, Madrid, Spain.
Félix
F
Pariente
Departamento Química Analítica y Análisis Instrumental, Universidad Autónoma de Madrid, 28049, Madrid, Spain; Instituto Madrileño de Estudios Avanzados (IMDEA) Nanociencia, Ciudad Universitaria de Cantoblanco, 28049, Madrid, Spain. Electronic address: encarnacion.lorenzo@uam.es.
Encarnación
E
Lorenzo
D001399:Azure Stains / Q000737:chemistry*; D004396:Coloring Agents / Q000737:chemistry*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics; D004247:DNA / Q000737:chemistry* / Q000235:genetics*; D004252:DNA Mutational Analysis / Q000379:methods*; D004563:Electrochemistry; D004566:Electrodes; D016480:Helicobacter pylori / Q000235:genetics; D008958:Models, Molecular; D009154:Mutation; D009690:Nucleic Acid Conformation; D009693:Nucleic Acid Hybridization; D010619:Phenazines / Q000737:chemistry*; D055672:Static Electricity; D013499:Surface Properties
D001399:Azure Stains; D004396:Coloring Agents; D010619:Phenazines; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D004247:DNA; C025818:Azure A; C009195:safranine T
DNA biosensor; Dye–DNA interaction; Gene mutation screening; Redox indicator