Enhancing single-cell transcriptomics using interposed anchor oligonucleotide sequences

Single-cell transcriptomics, which utilises barcodes and unique molecular identifiers (UMIs) for polyA+ mRNA capture, is compromised by oligonucleotide synthesis errors. To address this, we modified the oligonucleotide capture design and integrated an interposed anchor between the barcode and the UMI. This design significantly reduces the need to discard reads due to synthesis inaccuracies. Our results demonstrate that this anchor-enhanced design substantially improves gene expression profiles in droplet-based single-cell sequencing analyses.