Recursive construction of perfect DNA molecules and libraries from imperfect oligonucleotides

Methods Mol Biol. 2012:852:151-63. doi: 10.1007/978-1-61779-564-0_12.

Abstract

Making faultless complex objects from potentially faulty building blocks is a fundamental challenge in computer engineering, nanotechnology, and synthetic biology. We developed an error-correcting recursive construction procedure that attempts to address this challenge. Making DNA molecules from synthetic oligonucleotides using the procedure described here surpasses existing methods for de novo DNA synthesis in speed, precision, and amenability to automation. It provides for the first time a unified DNA construction platform for combining synthetic and natural DNA fragments, for constructing designer DNA libraries, and for making the faultless long synthetic DNA building blocks needed for de novo genome construction.

MeSH terms

  • Algorithms
  • Cloning, Molecular
  • DNA / biosynthesis*
  • DNA / genetics*
  • DNA / isolation & purification
  • DNA / metabolism
  • DNA Primers / genetics
  • DNA, Single-Stranded / metabolism
  • Exodeoxyribonucleases / metabolism
  • Gene Library*
  • Genetic Engineering / methods*
  • Oligodeoxyribonucleotides / metabolism*
  • Phosphorylation
  • Polymerase Chain Reaction
  • Synthetic Biology / methods

Substances

  • DNA Primers
  • DNA, Single-Stranded
  • Oligodeoxyribonucleotides
  • DNA
  • Exodeoxyribonucleases