Accurate gene synthesis with tag-directed retrieval of sequence-verified DNA molecules |
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Authors: | Jerrod J Schwartz Choli Lee Jay Shendure |
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Institution: | Department of Genome Sciences, University of Washington, Seattle, Washington, USA. |
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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. |
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