Direct Detection of Conserved Viral Sequences and Other Nucleic Acid Motifs with Solid-State Nanopores

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The fast and dependable recognition of nucleic acid sequences is important to a broad vary of fields together with genotyping, gene expression evaluation, and pathogen screening. For viral detection particularly, the aptitude is vital for optimum therapeutic response and stopping illness transmission. Right here, we report an strategy for detecting figuring out sequence motifs inside genome-scale single-strand DNA and RNA primarily based on solid-state nanopores. By designing DNA oligonucleotide probes with complementarity to focus on sequences inside a goal genome, we set up a protocol to yield affinity-tagged duplex molecules the identical size because the probe provided that the goal is current. The product can subsequently be certain to a protein chaperone and analyzed quantitatively with a selective solid-state nanopore assay. We first use a mannequin DNA genome (M13mp18) to validate the strategy, displaying the profitable isolation and detection of a number of goal sequences concurrently. We then show the protocol for the detection of RNA viruses by figuring out and concentrating on a extremely conserved sequence inside human immunodeficiency virus (HIV-1B).


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