BDB - Library Details

Library

Library Name: T7 LARFH gene-based phage display library (X3GX2)

Library ID:

483

Library Length:

Library Complexity:

7.40E+06

Library Titer:

Library From:

Akihiko Yamagishi (Tokyo University of Pharmacy and Life Sciences, Tokyo, Japan)

Library Randomness:

Semi-random

Library Scheme:

Library Topology:

Linear

Comments:

To create an LARFH gene library, amino acid residues at positions 21, 22, 23, 25, and 26 were subjected to semi-random mutagenesis. Gly24 was not mutated. Each of their codons was replaced with the NVS codon (N = A + T + G + C,V = A + G + C, S = G + C). Genes were then amplified by using two different primers so that the EcoRI and HindIII recognition sites could be added at the 5' and 3' termini, respectively. Amplified DNA fragments were digested with EcoRI and HindIII and then ligated to T7 Select Vector Arms (Merck Millipore, Germany). For in vitro packaging, ligated DNA fragments were added to T7 phage packaging extracts (Merck Millipore) and the mixtures were incubated at 25 °C for 2 h. After phage packaging, a T7 phage library containing 7.4e6 independent clones was constructed.

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