Earticle

초록

영어

We describe here ligation-based strategy to detect mutations in BRCA1 utilizing zip-code microarray technology. In our first approach, PCR was performed to amplify the genomic regions containing the mutation sites. The PCR products were then used as templates in a subsequent ligation reaction using two ligation primers that flanked the mutation site. The primary allele-specific primer is designed to contain respective zip-code at its 5/ end while the secondary primer is modified by biotin at its 3/ end. Then the nick between two ligation primers was sealed in presence of DNA ligase. The ligation products were then hybridized on the zip-code microarray followed by staining with streptavidine-cy3 to generate fluorescent signal. Using this strategy we successfully genotyped selected Korean-specificmutation sites of BRCA1 with a wild
type and two heterozygote mutant samples. Furthermore, we also demonstrated that ligase chain reaction using unamplified genomic DNA as direct templates is enough to generate sufficient signals for correct genotyping in a multiplexed manner, verifying first that PCR is not essential for this microarray-based strategy.

저자

• Agnishwar Girigoswami [ Department of Chemical and Biomolecular Engineering, Korea Advanced Institute of Science and Technology (KAIST) ]
• Cheulhee Jung [ Department of Chemical and Biomolecular Engineering, Korea Advanced Institute of Science and Technology (KAIST) ]
• Hyo Young Mun [ Department of Chemical and Biomolecular Engineering, Korea Advanced Institute of Science and Technology (KAIST) ]
• Hyun Gyu Park [ Department of Chemical and Biomolecular Engineering, Korea Advanced Institute of Science and Technology (KAIST) ]

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발행기관

간행물

표지보기 관심저널 등록

• 간행물명

  한국생물공학회 학술대회

• 간기

  반년간

• 수록기간

  1985~2013

• 십진분류

  KDC 476 DDC 576