Generating a synthetic genome by whole genome assembly: phiX174 bacteriophage from synthetic oligonucleotides.

@article{Smith2003GeneratingAS,
  title={Generating a synthetic genome by whole genome assembly: phiX174 bacteriophage from synthetic oligonucleotides.},
  author={Hamilton O. Smith and Clyde A. Hutchison and Cynthia M Pfannkoch and J. Craig Venter},
  journal={Proceedings of the National Academy of Sciences of the United States of America},
  year={2003},
  volume={100 26},
  pages={15440-5}
}
We have improved upon the methodology and dramatically shortened the time required for accurate assembly of 5- to 6-kb segments of DNA from synthetic oligonucleotides. As a test of this methodology, we have established conditions for the rapid (14-day) assembly of the complete infectious genome of bacteriophage X174 (5386 bp) from a single pool of chemically synthesized oligonucleotides. The procedure involves three key steps: (i). gel purification of pooled oligonucleotides to reduce… CONTINUE READING
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