×

Detection of nucleic acid sequence differences using the ligase detection reaction with addressable arrays

  • US 7,888,009 B2
  • Filed: 05/25/2004
  • Issued: 02/15/2011
  • Est. Priority Date: 02/09/1996
  • Status: Expired due to Fees
First Claim
Patent Images

1. A method for identifying a plurality of sequences differing by one or more single base changes, insertions, deletions, or translocations in a plurality of differing target nucleotide sequences comprising:

  • providing a sample potentially containing target nucleotide sequences differing by one or more single-base changes, insertions, deletions, or translocations;

    providing oligonucleotide probe sets, each set characterized by (a) a first oligonucleotide probe comprising a target-specific portion and (b) a second oligonucleotide probe comprising a target-specific portion and a further portion, wherein the further portion comprises a nucleic acid sequence of greater than 16 nucleotides that differs for each different target-specific portion, wherein the further portions of each set hybridize to their complementary portions under uniform hybridization conditions, wherein the nucleic acid sequence of a complement to one further portion differs from the nucleic acid sequence of a complement to another further portion, when aligned to each other, by at least 25% to minimize cross-hybridization of one further portion to a complementary portion of another further portion;

    providing a ligase;

    blending the sample, the oligonucleotide probe sets, and the ligase to form a ligation detection reaction mixture;

    subjecting the ligation detection reaction mixture to one or more ligation detection reaction cycles to form a ligated product sequence comprising (a) the target-specific portions and (b) the further portion if its target nucleotide sequence is present in the sample, wherein the further portion is comprised of a nucleotide sequence which is distinct from that of the target-specific portions to prevent hybridization between the target specific portions and the further portion; and

    detecting and distinguishing the presence of different target nucleotide sequences in the sample based on the further portion of the ligated product sequence.

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