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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.

SUBMITTER: Schwartz JJ

PROVIDER: S-EPMC3433648 | biostudies-literature | 2012 Sep

REPOSITORIES: biostudies-literature

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

Schwartz Jerrod J JJ Lee Choli C Shendure Jay J

Nature methods 20120812 9

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. ...[more]

PMID: 22886093

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