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Methods for the enzymatic assembly of polynucleotides and identification of polynucleotides having desired characteristics

  • US 6,635,453 B2
  • Filed: 05/10/2001
  • Issued: 10/21/2003
  • Est. Priority Date: 05/16/2000
  • Status: Expired due to Fees
First Claim
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1. A method of preparing a polynucleotide from a plurality of oligonucleotides, said method comprising:

  • a) blocking the 3′

    terminus of a first oligonucleotide with a blocking group to form a blocked oligonucleotide, wherein said first oligonucleotide comprises the 3′

    terminus of said polynucleotide;

    b) coupling the 3′

    terminus of a further oligonucleotide from said plurality of oligonucleotides to the 5′

    terminus of said blocked oligonucleotide to form a coupled oligonucleotide;

    wherein said coupling is carried out in the presence of an RNA ligase or a ribozyme;

    c) phosphorylating the 5′

    terminus of said further oligonucleotide;

    d) amplifying said coupled oligonucleotide to form an amplified oligonucleotide substantially free of blocking group; and

    e) repeating steps a) to d) with said amplified oligonucleotide until said polynucleotide is prepared.

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