title: Direct and nondestructive verification of PNA immobilization using click chemistry.
abstract: The fluorogenic 1,3-Huisgen dipolar cycloaddition reaction was used as part of a novel immobilization strategy of PNA capture probes on a microarray. By using this click chemistry, azidocoumarin-anchored PNA probes were immobilized on phenyl acetylene-modified glass slides with the simultaneous generation of the fluorescent triazolylcoumarin moiety. Since the emitting moieties are generated in the immobilization reaction itself, fluorescent signals can be used to directly monitor the integrity of immobilization in a nondestructive manner. By using this strategy, PNA microarrays were prepared and successfully employed to perform microarray-based diagnosis of selected mutations in the breast cancer susceptibility gene BRCA1.
authors.affiliation: Department of Chemical and Biomolecular Engineering (BK21 Program), Korea Advanced Institute of Science and Technology, 373-1 Guseong-dong, Yuseong-gu, Daejeon 305-701, Republic of Korea.
authors.forename: Seong Yoon
authors.initials: SY
authors.lastname: Lim
authors.forename: Won-Young
authors.initials: WY
authors.lastname: Chung
authors.forename: Hyeon Kyu
authors.initials: HK
authors.lastname: Lee
authors.forename: Myeong Soon
authors.initials: MS
authors.lastname: Park
authors.forename: Hyun Gyu
authors.initials: HG
authors.lastname: Park
mesh_terms: D000114:Acetylene / Q000031:analogs & derivatives / Q000737:chemistry; D001386:Azides / Q000737:chemistry; D001943:Breast Neoplasms / Q000175:diagnosis / Q000235:genetics; D003374:Coumarins / Q000737:chemistry; D004252:DNA Mutational Analysis / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry; D019398:Genes, BRCA1; D005898:Glass / Q000737:chemistry; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D015345:Oligonucleotide Probes / Q000737:chemistry; D020135:Peptide Nucleic Acids / Q000737:chemistry*; D013050:Spectrometry, Fluorescence / Q000379:methods*
chemical_list: D001386:Azides; D003374:Coumarins; D005456:Fluorescent Dyes; D015345:Oligonucleotide Probes; D020135:Peptide Nucleic Acids; C044736:phenylacetylene; C030123:coumarin; D000114:Acetylene