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- W4239441627 abstract "The ability of nucleic acids to form stable, sequence-specific complexes with foreign molecular probes has been exploited for a wide range of applications in life sciences, biotechnology, medicine, and forensics. Peptide nucleic acids (PNAs), nucleic acid analogs in which the negative sugar-phosphate backbone is replaced with a neutral peptide-like backbone, have been shown to display greater stability and sequence specificity to complementary ssDNA strands than natural DNA. This feature has been utilized in a number of applications in vitro and in vivo to ‘tag’ specific sequences. Moreover, the ability of PNA to locally displace one of the strands in double-stranded DNA (dsDNA), thus forming a P-loop, makes PNA an ideal candidate for dsDNA sequence detection. Here, we demonstrate a purely electrical detection method of short (8mer) sequences in dsDNA. Sub-5nm solid-state nanopores have recently demonstrated their capability in sizing DNA molecules as they translocate across the pore. Based on this finding, we show for the first time that short dsDNA sequences can be detected, label-free, on the single molecule level. We find that a ∼3.5 kbp long dsDNA ‘tagged’ with short PNA probes induces distinct secondary blockade levels in excess of those found on typical dsDNA molecules. Additionally, tagged molecules displayed significantly increased translocation times - and an increase in the distribution of those times. Furthermore, we demonstrate the ability to statistically discriminate between multi-tagged DNA and untagged DNA. We thus have established a foundation for the development of a radically new single-molecule platform for ultra-fast pathogen and mutation diagnostics1, ultimately impacting our ability to effectively respond to emerging infections or disease development on a personal level. 1) Singer, A., et al, (2009) “Nanopore-based sequence-specific detection of duplex DNA for genomic profiling”Journal of the American Chemical Society (under review)." @default.
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- W4239441627 date "2010-01-01" @default.
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- W4239441627 title "Nanopore-Based Sequence-Specific Detection of Duplex DNA for Genomic Profiling" @default.
- W4239441627 doi "https://doi.org/10.1016/j.bpj.2009.12.3258" @default.
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