We pinpoint PCR artifacts as the primary source of inaccurate quantification in both short- and long-read RNA sequencing, a problem that intensifies with an increase in PCR cycles in both bulk and single-cell sequencing contexts. To overcome this challenge, we engineered a novel unique molecular identifier (UMI) barcode composed of homotrimer nucleotide blocks. This design facilitates accurate quantification of RNA molecules, substantially improving molecular counting.
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This is a summary of: Sun, J. et al. Correcting PCR amplification errors in unique molecular identifiers to generate accurate numbers of sequencing molecules. Nat. Methods https://doi.org/10.1038/s41592-024-02168-y (2024)
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Homotrimer barcodes enable accurate counting of RNA molecules during high-throughput RNA sequencing. Nat Methods 21, 379–380 (2024). https://doi.org/10.1038/s41592-024-02169-x
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DOI: https://doi.org/10.1038/s41592-024-02169-x
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