Fig. 1

Workflow and performance of LAST-seq. a Schematic showing each step of the LAST-seq protocol. b Mapping rates of the LAST-seq reads to different genomic regions. The standard error of the mean (SEM) error bar is calculated from 10 cells. c Technical reproducibility of the LAST-seq data determined by the Pearson’s correlation coefficient (PCC) between two replicates using 15-pg HEK293T-extracted total RNA as the input. d Correlation between the number of UMIs and sequencing reads in the LAST-seq data of the ERCC RNA spike-in. e Correlation between the number of UMIs and the sequencing reads in the LAST-seq data of single HEK293T cells. f Linear correlation between the number of sequencing reads in the LAST-seq data and the input copy numbers of the ERCC RNA spike-in. g Linear correlation between the number of UMIs in the LAST-seq data and the input copy numbers of the ERCC RNA spike-in. h Coefficient of determination (R²) of the linear regression in (f) and (g), from 10 samples. The statistical analysis was performed by Welch t-test (*p ≤ 0.05). i Number of detected genes in single HEK293T cells with various sequencing depths. The SEM error bar is calculated from 10 and 9 cells for LAST-seq and SMART-seq, respectively