Array-based mutation detection of BRCA1 using direct probe/target hybridization

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We describe here an efficient micro array-based multiplex assay to detect Korean-specific mutations in breast cancer susceptibility gene BRCA1 using direct probe/target hybridization. Allele-specific oligonucleotides were covalently immobilized on an aldehyde-activated glass slide to prepare an oligonucleotide chip. From a wild-type sample, a two-step method was used to generate labeled multiplex polymerase chain reaction (PCR) amplification products of genomic regions containing the mutation sites. Amino allyl-dUTP, an amine-modified nucleotide, was incorporated during multiplex PCR amplifications and a monofunctional form of cyanine 3 dye was subsequently attached to the reactive amine group of the PCR products. Hybridization of the labeled PCR products to the oligonucleotide chip successfully identified all of the genotypes for the selected mutation sites. This work demonstrates that oligonucleotides chip-based analysis is a good candidate for efficient clinical testing for BRCA1 mutations when combined with the indirect strategy to prepare labeled target samples. (C) 2004 Elsevier Inc. All rights reserved.
Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
Issue Date
2005-02
Language
English
Article Type
Article
Keywords

OVARIAN-CANCER FAMILIES; GERMLINE MUTATIONS; RAPID DETECTION; BREAST; GENE

Citation

ANALYTICAL BIOCHEMISTRY, v.337, no.2, pp.332 - 337

ISSN
0003-2697
DOI
10.1016/j.ab.2004.11.034
URI
http://hdl.handle.net/10203/20356
Appears in Collection
CBE-Journal Papers(저널논문)
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