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LONG NUCLEIC ACID SEQUENCES CONTAINING VARIABLE REGIONS

  • US 20150159152A1
  • Filed: 12/09/2014
  • Published: 06/11/2015
  • Est. Priority Date: 12/09/2013
  • Status: Abandoned Application
First Claim
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1. A method of constructing a double stranded DNA fragment or library, said method comprising incorporating sequences between clonal or non-clonal double stranded DNA fragments (gene blocks), the method comprising:

  • a) forming a mixture comprised of a first gene block, a second gene block, and a bridging oligonucleotide set, said bridging oligonucleotide set comprising one or more bridging oligonucleotides, wherein each bridging oligonucleotide contains a first region that is hybridizable to a portion of the first gene block and a second region that is hybridizable to a portion of the second gene block;

    b) subjecting the mixture to reagents and conditions for PCR to assemble the gene blocks and bridge(s) thereby generating and optionally amplifying a double stranded DNA fragment or library, wherein the sequence generated is comprised of the first gene block, a bridge sequence of the bridging oligonucleotide(s), if any, that did not hybridize to a gene block, and the second gene block.

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