Here the recognition of a single‐point mutation in oligonucleotides is described by using nanopore measurements. The translocation behavior of a series of mutated DNA strands, hybridized with a complementary DNA probe, is analyzed via blocking current and unzipping time. Discernment of the mutation position at the single nucleotide level is achieved by analysis of a 2D graph of the bootstrapped translocation data. The proposed approach provides a useful tool for the mutation detection of oligonucleotides secreted from tumor cells and is applicable in simple and label‐free diagnoses as a nanopore liquid biopsy.
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