BDB - Library Details

Library

Library Name: PL12 yeast display library (X12)

Library ID:

333

Library Length:

Library Complexity:

Library Titer:

Library From:

Angela M. Belcher

Library Randomness:

Completely random

Library Scheme:

NNK

Library Topology:

Linear

Comments:

A yeast display peptide library was created from a degenerate oligonucleotide pool encoding random 12-mer peptides with a NNK bias. Peptides were displayed as fusions to the C-terminus of Aga2, which is encoded on a plasmid downstream of a Gal-based promoter. The expression vectors were maintained in S. cerevisiae strain EBY100, which has Aga1 under control of a Gal-based promoter integrated in its genome, Oligonucleotides used in library construction were obtained from Oligos, Etc. The degenerate oligonucleotide pool encoding the random 12-mer peptide library (PL12LibrBstXI) 50-pGTGGCGGTAGCGGC-(NNK)12-TAGCTAGCTAGGCCAGTAGC, synthesized with an NNK bias in order to reduce the stop codon frequency, was annealed to two short primers (5'BstXIanneal) 5'-pGCCGCTACCGCCACCGCC and (3'BstXIanneal) 5'-pCTGGCCTAGCTAGCTA. The resulting hybrid DNA library containing duplexed BstXI sticky ends was directly ligated into the BstXI sites of pBPZ, and electroporated into E. coli Electromax DH10B (Invitrogen, Carlsbad, CA) for amplification on plated LB Ampt media.

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