Detection of nucleic acid sequence differences using coupled ligase detection and polymerase chain reactions

CAFC
  • US 7,332,285 B2
  • Filed: 10/31/2006
  • Issued: 02/19/2008
  • Est. Priority Date: 05/29/1996
  • Status: Expired due to Fees
First Claim
Patent Images

1. A method for identifying one or more different target nucleotide sequences comprising:

  • providing a sample potentially containing one or more target nucleotide sequences comprising sequence differences;

    performing a ligase detection reaction to form a ligation product comprising a first target specific portion complementary to the target nucleotide sequence and a second target specific portion complementary to the target nucleotide sequence, wherein the ligation product further comprises a 5′

    upstream primer-specific portion and a 3′

    downstream primer-specific portion, wherein the first target specific portion and the second target specific portion are located between the 3′

    downstream primer-specific portion and the 5′

    upstream primer-specific portion;

    providing an oligonucleotide primer set comprising (a) an upstream primer containing the same sequence as the 5′

    upstream primer-specific portion of the ligation product sequence and (b) a downstream primer complementary to the 3′

    downstream primer-specific portion of the ligation product sequence;

    performing a polymerase chain reaction to form extension products comprising the ligation product sequence and/or complements thereof; and

    ,detecting the extension products to identify one or more target nucleotide sequences in the sample.

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