title: Direct covalent attachment of DNA microarrays by rapid thiol-ene "click" chemistry.
abstract: A rapid strategy for the covalent immobilization of DNA onto silicon-based materials using the UV-initiated radical thiol-ene reaction is presented in this study. Following this approach, thiol- and alkene-modified oligonucleotide probes were covalently attached in microarray format, reaching immobilization densities around 6 pmol·cm(-2). The developed methodology presents the advantages of spatially controlled probe anchoring (using a photomask), direct attachment without using cross-linkers (one-pot fashion), and short irradiation times (20 min). Using the described strategy, hybridization efficiencies up to 65% with full complementary strands were reached. The approach was evaluated by scoring single-base pair mismatches with discrimination ratios around 15. Moreover, the efficacy of the proposed DNA detection scheme is further demonstrated through the assay on a genomic target of bacterial Escherichia coli.
authors.affiliation: Centro de Reconocimiento Molecular y Desarrollo Tecnológico, Departamento de Química, Universitat Politècnica de València , Camino de Vera s/n, 46022 Valencia, Spain.
authors.forename: Jorge
authors.initials: J
authors.lastname: Escorihuela
authors.forename: María-José
authors.initials: MJ
authors.lastname: Bañuls
authors.forename: Santiago
authors.initials: S
authors.lastname: Grijalvo
authors.forename: Ramón
authors.initials: R
authors.lastname: Eritja
authors.forename: Rosa
authors.initials: R
authors.lastname: Puchades
authors.forename: Angel
authors.initials: A
authors.lastname: Maquieira
mesh_terms: D000475:Alkenes / Q000737:chemistry*; D057930:Click Chemistry; D004247:DNA / Q000032:analysis*; D004926:Escherichia coli / Q000302:isolation & purification; D005453:Fluorescence; D020411:Oligonucleotide Array Sequence Analysis; D015345:Oligonucleotide Probes / Q000737:chemistry*; D017646:Organosilicon Compounds / Q000737:chemistry; D010316:Particle Size; D013438:Sulfhydryl Compounds / Q000737:chemistry*; D013499:Surface Properties; D014466:Ultraviolet Rays
chemical_list: D000475:Alkenes; D015345:Oligonucleotide Probes; D017646:Organosilicon Compounds; D013438:Sulfhydryl Compounds; D004247:DNA