Figure 2.

Strategies to analyze phage library by second- and third-generation sequencing. On the left, the workflow for extraction of variable fragments compatible with second-generation sequencing platforms (e.g., Illumina MiSeq) is shown. Full-length scFvs and VH fragments are extracted from dsDNA of phages from E. coli. On the right, the workflow for the application of third-generation sequencing of entire scFvs by circular consensus sequencing is represented. Molecular barcoding can be included in both strategies for multiplexing samples. The indicated pros (green font) and cons (red font) are referred to as the current state-of-the-art.