Direct and nondestructive verification of PNA immobilization using click chemistry.
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.
Lim
Seong Yoon
SY
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.
Chung
Won-Young
WY
Lee
Hyeon Kyu
HK
Park
Myeong Soon
MS
Park
Hyun Gyu
HG
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*
D001386:Azides; D003374:Coumarins; D005456:Fluorescent Dyes; D015345:Oligonucleotide Probes; D020135:Peptide Nucleic Acids; C044736:phenylacetylene; C030123:coumarin; D000114:Acetylene