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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.

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