Nanopore sequencing of single-cell transcriptomes with scCOLOR-seq
Martin Philpott, Jonathan Watson, Anjan Thakurta, Tom Brown, Tom Brown, Udo Oppermann, Adam P. Cribbs
Abstract
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.
Topics & Concepts
BarcodeNanopore sequencingComputational biologyComplementary DNAIdentifierOligonucleotideSingle cell sequencingNanoporeMinionDNA sequencingGene isoformBiologyDNAComputer scienceGeneGeneticsExome sequencingNanotechnologyPhenotypeOperating systemMaterials scienceProgramming languageSingle-cell and spatial transcriptomicsCancer Genomics and DiagnosticsAdvanced biosensing and bioanalysis techniques