Accurate gene synthesis with tag-directed retrieval of sequence-verified DNA molecules

Nat Methods. 2012 Sep;9(9):913-5. doi: 10.1038/nmeth.2137. Epub 2012 Aug 12.

Abstract

We present dial-out PCR, a highly parallel method for retrieving accurate DNA molecules for gene synthesis. A complex library of DNA molecules is modified with unique flanking tags before massively parallel sequencing. Tag-directed primers then enable the retrieval of molecules with desired sequences by PCR. Dial-out PCR enables multiplex in vitro clone screening and is a compelling alternative to in vivo cloning and Sanger sequencing for accurate gene synthesis.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA / biosynthesis
  • DNA / genetics*
  • Escherichia coli / genetics
  • Genes / genetics*
  • Multiplex Polymerase Chain Reaction / methods*
  • Oligonucleotide Array Sequence Analysis*
  • Oligonucleotides / genetics

Substances

  • Oligonucleotides
  • DNA