title: Accurate gene synthesis with tag-directed retrieval of sequence-verified DNA molecules.
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.
authors.affiliation: Department of Genome Sciences, University of Washington, Seattle, Washington, USA.
authors.forename: Jerrod J
authors.initials: JJ
authors.lastname: Schwartz
authors.forename: Choli
authors.initials: C
authors.lastname: Lee
authors.forename: Jay
authors.initials: J
authors.lastname: Shendure
mesh_terms: D004247:DNA / Q000096:biosynthesis / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D005796:Genes / Q000235:genetics*; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis; D009841:Oligonucleotides / Q000235:genetics
chemical_list: D009841:Oligonucleotides; D004247:DNA