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Direct microRNA Sequencing Using Nanopore-Induced Phase-Shift Sequencing

iScience. 2020; 
Zhang J, Yan S, Chang L, Guo W, Wang Y, Wang Y, Zhang P, Chen HY, Huang S.
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Custom DNA/RNA Oligos All HPLC-purified DNA oligonucleotides, including DNA-miRNA template strands, primer, blocker, DNA linker and miR-21 (Table S1) were custom synthesized by Genscript (New Jersey, USA). Get A Quote

Abstract

MicroRNAs (miRNAs) are a class of short non-coding RNAs that function in RNA silencing and post-transcriptional gene regulation. However, direct characterization of miRNA is challenging due to its unique properties such as its low abundance, sequence similarities, and short length. Although urgently needed, single molecule sequencing of miRNA has never been demonstrated, to the best of our knowledge. Nanopore-induced phase-shift sequencing (NIPSS), which is a variant form of nanopore sequencing, could directly sequence any short analytes including miRNA. In practice, NIPSS clearly discriminates between different identities, isoforms, and epigenetic variants of model miRNA sequences. This work thus demonstrates ... More

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