BDB - Library Details

Library

Library Name: X6 fdMED1 phage display library

Library ID:

349

Library Length:

Library Complexity:

Library Titer:

Library From:

Jean-Pierre Mach

Library Randomness:

Completely random

Library Scheme:

NNK

Library Topology:

Linear

Comments:

The vector fdMED1 was used for the library construction. In order to avoid the background production of wild-type M13 phage during the selections, a new polylinker cloning site containing several stop codons was constructed between the two unique restriction sites ApaLI and NotI of the original fdDOG1 vector. The construction of this new vector (fdMED1) was performed as follows: the peptide leader sequence (LpelB) of the vector backbone pHEN1 was amplified by PCR with Taq polymerase (Perkin Elmer) using oligo-1 and oligo-fdSEQ01FOR. The reaction mixture (50 mL) was cycled 30 times. The resulting DNA fragment was subcloned into ApaLI and NotI digested fdDOG1 and used to transform Escherichia coli TG1. Clones containing the correct insert were identified by DNA sequencing. In addition, a multiple cloning site (MCS) was generated by inserting annealed oligonucleotides (oligo-4 and oligo-5) into SfiI/NotI digested vector. The DNA was electroporated in E. coli MC1061, thus generating a new fd vector called fdMED1.

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