PLoS ONE (Jan 2017)

QuickLib, a method for building fully synthetic plasmid libraries by seamless cloning of degenerate oligonucleotides.

  • Pierre Galka,
  • Elisabeth Jamez,
  • Gilles Joachim,
  • Patrice Soumillion

DOI
https://doi.org/10.1371/journal.pone.0175146
Journal volume & issue
Vol. 12, no. 4
p. e0175146

Abstract

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Incorporation of synthetic degenerate oligonucleotides into plasmids for building highly diverse genetic libraries requires efficient and quantitative DNA manipulation. We present a fast and seamless method for generating libraries of PCR-synthesized plasmids designed with a degenerate sequence and short overlapping ends. Our method called QuickLib should find many applications in synthetic biology; as an example, we easily prepared genetic libraries of Escherichia coli expressing billions of different backbone cyclic peptides.