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Enzymatic synthesis of repeat regions of oligonucleotides

  • US 5,516,664 A
  • Filed: 12/02/1993
  • Issued: 05/14/1996
  • Est. Priority Date: 12/23/1992
  • Status: Expired due to Fees
First Claim
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1. A method for synthesizing a repeat region of an oligonucleotide having a defined sequence, said repeat region including a repeated nucleotide that appears more than once in succession, comprising the steps of:

  • (a) enzymatically coupling an oligonucleotide primer with a 3'"'"'-phosphate-blocked repeated nucleotide to form a 3'"'"'-phosphate blocked primer;

    (b) removing the 3'"'"'-phosphate blocking group from the 3'"'"'-phosphate-blocked primer using a 3'"'"'-phosphatase enzyme substantially without removing the 3'"'"'-phosphate blocking group from unreacted 3'"'"'-phosphate-blocked repeated nucleotide; and

    (c) repeating steps (a) and (b) using unreacted 3'"'"'-phosphate-blocked repeated nucleotide from step (b) as the 3'"'"'-phosphate-blocked repeated nucleotide of step (a) and the deblocked primer product of step (b) as the oligonucleotide primer of step (a) without prior separation of the unreacted 3'"'"'-phosphate-blocked repeated nucleotide from the deblocked primer product.

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