Fig. 3

Prime-seq has similar sensitivity and power compared to TruSeq (MAQC-III data). A Mapped reads, UMIs (dashed line, only prime-seq), and B detected genes (exonic + intronic reads) at varying sequencing depths between TruSeq data from the MAQC-III Study and matched prime-seq data, show prime-seq and TruSeq are similarly sensitive (filtering parameters: detected UMI ≥ 1, detected gene present in at least 25% of samples and is protein coding). C Accuracy, measured by spike-in molecules, is similarly high in both methods (R² = 0.94). D The distribution of genes across mean expression is similar for both methods, as well as the dispersion, which follows a Poisson distribution (dark gray dashed line) for lower expressed genes and then increases as technical variation increases for highly expressed genes. The local polynomial regression fit between mean and dispersion estimates per method is shown in solid lines with 95% variability band per gene shown in dashed lines. E Power analysis at a sequencing depth of 10 million reads shows almost identical power between prime-seq and TruSeq, and a similar increase at varying sample size for F mean expression and G absolute log2 fold change. Data filtering parameters: detected UMI ≥ 1, detected gene present in at least 25% of samples