Microarray-based detection of Korean-specific BRCA1 mutations.
A reliable multiplex assay procedure to detect human genetic mutations in the breast cancer susceptibility gene BRCA1 using zip-code microarrays and single base extension (SBE) reactions is described. Multiplex PCR amplification was performed to amplify the genomic regions containing the mutation sites. The PCR products were then employed as templates in subsequent multiplex SBE reactions using bifunctional primers carrying a unique complementary zip sequence in addition to a mutation-site-specific sequence. The SBE primers, terminating one base before their mutation sites, were extended by a single base at a mutation site with a corresponding biotin-labeled ddNTP. Hybridization of the SBE products to zip-code microarrays was followed by staining with streptavidin-Cy3, leading to successful genotyping of several selected BRCA1 mutation sites with wild-type and heterozygote mutant samples from breast cancer patients. This work has led to the development of a reliable DNA microarray-based system for the diagnosis of human genetic mutations.
Jung
Cheulhee
C
Department of Chemical and Biomolecular Engineering, KAIST, 373-1 Guseong-dong, Yuseong-gu, Daejeon, 305-701, Republic of Korea.
Yim
Seong-Chun
SC
Cho
Dae-Yeon
DY
Chang
Ho Nam
HN
Park
Hyun Gyu
HG
D019313:BRCA1 Protein / Q000235:genetics*; D001943:Breast Neoplasms / Q000235:genetics*; D005260:Female; D006580:Genetic Carrier Screening; D005820:Genetic Testing / Q000379:methods; D005838:Genotype; D006801:Humans; D007723:Korea; D009154:Mutation; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation / Q000379:methods
D019313:BRCA1 Protein