earticle

영어

We developed new strategy for the detection of mutations in breast cancer susceptibility gene BRCA on PNA zip-code microarray by Single strand specific (SSS) nuclease. SSS nucleases was reported that is working from different sources to cleave single base pair mismatches in heteroduplex DNA templates used for mutation and single nucleotide polymorphism analysis. We focused this property and then we combined to detect of BRCA mutation site using on PNA zip-code microarray by SSS nuclease. The PCR products were performed to amplify the genomic regions containing the mutation sites. The PCR products were then employed as templates in a subsequent SSS nuclease reaction using chimeric primers with biotin-labeled 3' complementarity to the specific mutation site and 5' complementarity to the respective PNA zip-code sequence on the microarray. Hybridization of the labeled primers to the PCR products and This primer was cleaved by sss nuclease at each wild site in the presence of biotin-labeled region and mutation site in the absent of biotin-labeled region, and the products were hybridized to the PNA zip-code microarray. We conclude that SSS nuclease assay on PNA zip-code microarray can be effectively used for high-throughput detection of mutation.