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Single Nucleotide Discrimination by α-Hemolysin Nanopore

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We in this paper presented a single-base resolution method for discrimination of single base pair differences within DNA duplexes containing A-T matched base pairs and A-C mismatched base pairs using α-hemolysin (α-HL) protein nanopore embedded in a lipid bilayer. A 17-mer probe respectively annealed to two 57-mer target ssDNA which had poly(dT)20 tails on both 5' and 3' ends, forming single base-pair differentiating DNA duplexes containing the matched A-T base pairs and A-C mismatched base pairs which were then voltage-driven captured and stacked into the α-HL constriction zone under a biased potential. After a long unzipping process, the longer stranded and probe DNA was successively separated and translocated through the α-HL channel, then generating a characteristic signal. Ion channel recordings were used to detect and compare the unzipping process. The duplex structures exhibited different unzipping time duration under applied voltage by firstly threading either the 3' or 5' overhangs into the α-hemolysin channel that permitted easy discrimination between the completely matched and single-mismatched base pairs. The simple biosensing strategy will open up a variety of applications on SNP sites detection, diseases classification, medical screening and diagnosis.
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Keywords: DNA UNZIPPING; NANOPORE; SINGLE NUCLEOTIDE DISCRIMINATION

Document Type: Short Communication

Publication date: August 1, 2016

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  • Nanoscience and Nanotechnology Letters (NNL) is a multidisciplinary peer-reviewed journal consolidating nanoscale research activities in all disciplines of science, engineering and medicine into a single and unique reference source. NNL provides the means for scientists, engineers, medical experts and technocrats to publish original short research articles as communications/letters of important new scientific and technological findings, encompassing the fundamental and applied research in all disciplines of the physical sciences, engineering and medicine.
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