Gold nanoparticle-based label-free detection of BRCA1 mutations utilizing DNA ligation on DNA microarray.
Here we describe a label-free detection strategy for point mutation in breast cancer susceptibility gene BRCA1 utilizing ligation chain reaction (LCR) and zip-code array. We amplified the genomic regions containing mutation sites by polymerase chain reaction (PCR) to prepare the template for LCR. Then we ligated a primary probe extended by zip-code complementary at the 5' end with a 3'-thiol modified secondary probe. The resulting ligated product was labeled with gold nanoparticles and allowed to hybridize with corresponding zip-code sequences on the microarray. Finally, we applied silver enhancement to amplify the signals. Using this strategy, we successfully genotyped BRCA1 mutation, which was detected by naked eye.
Department of Chemical and Biomolecular Engineering, Korea Advanced Institute of Science and Technology, 373-1 Guseong-dong, Yuseong-gu, Daejeon, 305-701, Republic of Korea.
Agnishwar
A
Girigoswami
Taihua
T
Li
Cheulhee
C
Jung
Hyo Young
HY
Mun
Hyun Gyu
HG
Park
D001483:Base Sequence; D004247:DNA / Q000235:genetics*; D017931:DNA Primers; D019398:Genes, BRCA1; D006046:Gold / Q000737:chemistry*; D053768:Metal Nanoparticles; D009154:Mutation; D020411:Oligonucleotide Array Sequence Analysis; D016133:Polymerase Chain Reaction
D017931:DNA Primers; D006046:Gold; D004247:DNA