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Detection of Large Deletion in Human Brca1 Gene in Human Breast Carcinoma Mcf-7 Cells by Using Dna-Silver Nanoclusters Publisher Pubmed



Borghei YS1 ; Hosseini M1, 2 ; Ganjali MR3, 4
Authors
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Authors Affiliations
  1. 1. Department of Life Science Engineering, Faculty of New Sciences and Technologies, University of Tehran, Tehran, Iran
  2. 2. Medical Biomaterials Research Center, Tehran University of Medical Sciences, Tehran, Iran
  3. 3. Center of Excellence in Electrochemistry, Faculty of Chemistry, University of Tehran, Tehran, Iran
  4. 4. Biosensor Research Center, Endocrinology and Metabolism Molecular-Cellular Sciences Institute, Tehran University of Medical Sciences, Tehran, Iran

Source: Methods and Applications in Fluorescence Published:2018


Abstract

Here we describe a label-free detection strategy for large deletion mutation in breast cancer (BC) related gene BRCA1 based on a DNA-silver nanocluster (NC) fluorescence upon recognition-induced hybridization. The specific hybridization of DNA templated silver NCs fluorescent probe to target DNAs can act as effective templates for enhancement of AgNCs fluorescence, which can be used to distinguish the deletion of BRCAl due to different fluorescence intensities. Under the optimal conditions, the fluorescence intensity of the DNA-AgNCs at emission peaks around 440 nm (upon excitation at 350 nm) increased with the increasing deletion type within a dynamic range from 1.0 × 10-10 to 2.4 × 10-6 M with a detection limit (LOD) of 6.4 × 10-11 M. In this sensing system, the normal type shows no significant fluorescence; on the other hand, the deletion type emits higher fluorescence than normal type. Using this nanobiosensor, we successfully determined mutation using the non-amplified genomic DNAs that were isolated from the BC cell line. © 2017 IOP Publishing Ltd.
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