使用scCOLOR-seq对单细胞转录组进行纳米孔测序。

Philpott Martin, Watson Jonathan, Thakurta Anjan, Brown Tom, Brown Tom, Oppermann Udo, Cribbs Adam P

Botnar Research Centre, Nuffield Department of Orthopedics, Rheumatology and Musculoskeletal Sciences, National Institute of Health Research Oxford Biomedical Research Unit (BRU), University of Oxford, Oxford, UK.

Oxford Centre for Translational Myeloma Research University of Oxford, Oxford, UK.

Nat Biotechnol. 2021 Dec;39(12):1517-1520. doi: 10.1038/s41587-021-00965-w. Epub 2021 Jul 1.

Here we describe single-cell corrected long-read sequencing (scCOLOR-seq), which enables error correction of barcode and unique molecular identifier oligonucleotide sequences and permits standalone cDNA nanopore sequencing of single cells. Barcodes and unique molecular identifiers are synthesized using dimeric nucleotide building blocks that allow error detection. We illustrate the use of the method for evaluating barcode assignment accuracy, differential isoform usage in myeloma cell lines, and fusion transcript detection in a sarcoma cell line.

在此，我们描述了单细胞校正长读长测序（scCOLOR-seq），它能够对条形码和独特分子标识符寡核苷酸序列进行纠错，并允许对单细胞进行独立的cDNA纳米孔测序。条形码和独特分子标识符是使用允许错误检测的二聚体核苷酸构建块合成的。我们展示了该方法在评估条形码分配准确性、骨髓瘤细胞系中异构体差异使用以及肉瘤细胞系中融合转录本检测方面的应用。