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• W2893961808 abstract "Although single-cell mRNA sequencing has been a powerful tool to explore cellular heterogeneity, the sequencing of small RNA at the single-cell level (sc-sRNA-seq) remains a challenge, as these have no consensus sequence, are relatively low abundant, and are difficult to amplify in a bias-free fashion. We present two methods of single-cell-lysis that enable sc-sRNA-seq. The first method is a chemical-based technique with overnight freezing while the second method leverages on-chip electrical lysis of plasma membrane and physical extraction and separation of cytoplasmic RNA via isotachophoresis. We coupled these two methods with off-chip small RNA library preparation using CleanTag modified adapters to prevent the formation of adapter dimers. We then demonstrated sc-sRNA-seq with single K562 human leukemic cells. Our approaches offer a relatively short hands-on time of 6 h and efficient generation of on-target reads. The sc-sRNA-seq with our approaches showed detection of miRNA with various abundances ranging from 16 000 copies/cell to about 10 copies/cell. We anticipate this approach will create a new opportunity to explore cellular heterogeneity through small RNA expression." @default.
• W2893961808 created "2018-10-05" @default.
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• W2893961808 date "2018-09-27" @default.
• W2893961808 modified "2023-10-16" @default.
• W2893961808 title "Efficient Production of On-Target Reads for Small RNA Sequencing of Single Cells Using Modified Adapters" @default.
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• W2893961808 doi "https://doi.org/10.1021/acs.analchem.8b02773" @default.
• W2893961808 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/6233959" @default.
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• W2893961808 hasPublicationYear "2018" @default.
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