×

Method for enzymatic synthesis of oligonucleotides

  • US 5,602,000 A
  • Filed: 06/23/1995
  • Issued: 02/11/1997
  • Est. Priority Date: 12/23/1992
  • Status: Expired due to Fees
First Claim
Patent Images

1. A method for synthesizing an oligonueleotide of a defined sequence, comprising the steps of:

  • (a) combining (1) an oligonucleotide primer and (2) a blocked nucleotide or a blocked nucleotide precursor that forms a blocked nucleotide in situ, in a reaction mixture in the presence of a chain extending enzyme effective to couple the blocked nucleotide to the 3'"'"'-end of the oligonucleotide primer such that a primer-blocked nucleotide product is formed, wherein the blocked nucleotide comprises a nucleotide to be added to form part of the defined sequence and a blocking group attached to the 3'"'"'-end of the nucleotide effective to prevent the addition of more than one blocked nucleotide to the primer;

    (b) removing the blocking group from the 3'"'"'-end of the primer-blocked nucleotide product to form a primer-nucleotide product, whereby the reaction mixture contains any unreacted starting materials that may remain, primer-nucleotide product and reaction by-products; and

    (c) repeating at least one cycle of steps (a) and (b) using the primer-nucleotide product from step (b) as the oligonucleotide primer of step (a) in the subsequent cycle without separation of the primer-nucleotide product from the remainder of the reaction mixture.

View all claims
  • 0 Assignments
Timeline View
Assignment View
    ×
    ×