Creation of libraries with long ORFs by polymerization of a microgene.

@article{Shiba1997CreationOL,
  title={Creation of libraries with long ORFs by polymerization of a microgene.},
  author={Kiyotaka Shiba and Yoshinori Takahashi and Tetsuo Noda},
  journal={Proceedings of the National Academy of Sciences of the United States of America},
  year={1997},
  volume={94 8},
  pages={
          3805-10
        }
}
We describe a novel method for constructing pools of DNA sequences that encode large proteins with molecular diversity. Sets of primer pairs that form 8 to 10 complementary base pairs in the 3' region and have double mismatch pairs at their 3'-OH ends were designed so that primer dimers recreated short stretches of DNA (microgenes) devoid of termination codons. Cycles of denaturation and elongation reactions with a pair of primers, four dNTPs, and 3'-5' exo+ thermostable DNA polymerase gave… CONTINUE READING
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