Next, we repeated the PASSPORT-seq assay again with another five biological replicates in each of the three cell lines to validate the observed results. A separate cDNA and sequencing library was created for the experiments. A strong correlation (R2 = 0.98; p < 0.05) was observed in the results from the plasmid pool from the first sequencing run and those from the second sequencing run (see Supplementary Figure 5). There was a high correlation (R2 = 0.74; p < 0.05) in the results between the first and second set of biological replicates (Figure 2 and Supplementary Figure 6). This strong correlation between results of the first sequencing run with those of the second sequencing run across the three cell lines demonstrates the high reproducibility of the observed results.
