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After incubation at 37[degrees]C for 15 min, phagemids were titrated with 1 x NZY broth.
Briefly, phage were propagated by infection of Escherichia coli K91 B1ueKan cells (27), followed by growth of the infected cells for 16 h in NZY medium containing 20 mg/L tetracycline (28).
Aliquots of the constructed leaf roll library were plated out onto solid NZY medium and single plaques randomly picked and stored in SM buffer [100 mM NaCl, 8 mM Mg[SO.sub.4] 7[H.sub.2]O, 20 mM Tris-HCl pH 7.5, 0.01% (w/v) gelatin] at 4 [degrees] C.
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