Microarray-based detection of Korean-specific BRCA1 mutations

Cheulhee Jung, Seong Chun Yim, Dae Yeon Cho, Ho Nam Chang, Hyun Gyu Park

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)405-413
Number of pages9
JournalAnalytical and Bioanalytical Chemistry
Volume391
Issue number1
DOIs
Publication statusPublished - 2008 May 1

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Keywords

  • BRCA
  • DNA chip
  • Mutation detection
  • Single base extension
  • Zip-code microarray

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry

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