| Description: | First, three rounds of serial amplification of the phage-displayed peptide library without any panning steps (i.e., no exposure to a target) allowed fast-propagating clones to be enriched in the amplified pool. The next step took advantage of the fact that faster phage have significantly higher concentrations 135 minutes into the infection of E. coli cells. The serially amplified library was plated, and randomly selected plaques were used to infect separate 1 mL cultures of E. coli. At 135 minutes, the cultures were diluted appropriately and plated. Ph-VTAHGGR (G6813A) was identified to have relatively high titers (e.g., >100 plaques when 10 μL of 106x diluted culture was plated), where Ph- represents the phage, the displayed 7-mer peptide follows, and the gene II 5'-UTR mutation is in parentheses. |
