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Strategy of the library construction and panning. (A) Flow of the project, starting from B-cell collection, PCR amplification of the VNAR sequences, assemblage of VNAR fragments to vector backbone, electroporation of TG1 cells to make the library (containing 1.2 × 1010 individual clones), and panning on plastic plates coated with different antigens. (B) Diagramed protocol for the assemblage of VNAR fragments with vector backbone by PCR-EASeL.
