×

Combinatorial selection of oligonucleotide aptamers

  • US 20030027184A1
  • Filed: 04/11/2002
  • Published: 02/06/2003
  • Est. Priority Date: 10/26/1998
  • Status: Active Grant
First Claim
Patent Images

1. A method of identifying an aptamer comprising the steps of:

  • (a) synthesizing a random phosphodiester oligonucleotide combinatorial library wherein constituent oligonucleotides comprise at least a set of 5′ and

    /or 3′

    PCR primer nucleotide sequences flanking a randomized nucleotide sequence;

    (b) amplifying the library enzymatically using a mix of four nucleotides, wherein at least a percentage of one of the nucleotides is modified, to form a modified oligonucleotide combinatorial library;

    (c) contacting the modified oligonucleotide combinatorial library with a target molecule and isolating a subset of oligonucleotides binding to the target molecule;

    (e) amplifying the subset of binding oligonucleotides enzymatically using a mix of four nucleotide substrates, wherein at least one nucleotide substrate is modified, to form a modified oligonucleotide sub-library; and

    (f) repeating steps (c)-(e) iteratively until at least one aptamer comprising a modified oligonucleotide population of defined sequence is obtained.

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