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.
Girigoswami
Agnishwar
A
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.
Li
Taihua
T
Jung
Cheulhee
C
Mun
Hyo Young
HY
Park
Hyun Gyu
HG
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