In vitro recombination method

Young, Lei; Smith, Hamilton O.; Gibson, Daniel Glenn

Title: In vitro recombination method

Patent · Tue Jan 03 00:00:00 EST 2017

OSTI ID:1531965

Young, Lei; Smith, Hamilton O.; Gibson, Daniel Glenn

The present invention relates, e.g., to in vitro method, using isolated protein reagents, for joining two double stranded (ds) DNA molecules of interest, wherein the distal region of the first DNA molecule and the proximal region of the second DNA molecule share a region of sequence identity, comprising contacting the two DNA molecules in a reaction mixture with (a) a non-processive 5′ exonuclease; (b) a single stranded DNA binding protein (SSB) which accelerates nucleic acid annealing; (c) a non strand-displacing DNA polymerase; and (d) a ligase, under conditions effective to join the two DNA molecules to form an intact double stranded DNA molecule, in which a single copy of the region of sequence identity is retained. The method allows the joining of a number of DNA fragments, in a predetermined order and orientation, without the use of restriction enzymes.

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Research Organization:: Synthetic Genomics, Inc., La Jolla, CA (United States)

Sponsoring Organization:: USDOE

DOE Contract Number:: FG02-02ER63453

Assignee:: Synthetic Genomics, Inc. (La Jolla, CA)

Patent Number(s):: 9,534,251

Application Number:: 12/750,571

OSTI ID:: 1531965

Resource Relation:: Patent File Date: 2010-03-30

Country of Publication:: United States

Language:: English

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Efficient and rapid method for assembling and cloning double-stranded DNA fragments Rose, Scott; Walder, Joseph A.; Beltz, Kristin https://doi.org/https://image-ppubs.uspto.gov/dirsearch-public/print/downloadPdf/9951327 US Patent Document 9,951,327	patent	April 2018

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