Figure 4: BC32 construct.
From: Limitations and challenges of genetic barcode quantification
(a) The LeGO-vectors were additionally equipped with the second Illumina adaptor and dedicated restriction enzyme recognition sites. The barcodes consist of 32 random nucleotides separated by fixed triplets. (b) Overview of the minimal HD of the picked barcodes. (c) Quantitative ddPCR based on integration sites was performed directly after generating the miniBulks. (d) The frequency plot of one particular miniBulk depicts all identified barcodes with more than two reads (log scale). Every barcode is colored according to its minimal HD to one of the five original barcodes. The original four barcodes are depicted in the inset. (e) The average abundance of all four original barcodes over all miniBulk replicates after 40 cycles of PCR (standard protocol, gDNA). (f) Sequence similarities are visualized with a network based graph (ripple plot). Barcodes are represented as nodes, sequence similarities of a HD = 1 between two barcodes are visualized as links and node sizes reflect read counts. The minimal HD of each barcode to one of the original barcodes is color-coded according to the depicted legend.
