DETECTION OF NUCLEIC ACID SEQUENCE DIFFERENCES USING COUPLED LIGASE DETECTION AND POLYMERASE CHAIN REACTIONS

摘要

The present invention relates to a method for identifying a target nucleotide sequence. This method involves forming a ligation product on a target nucleotide sequence in a ligation detection reaction mixture, amplifying the ligation product to form an amplified ligation product in a polymerase chain reaction (PCR) mixture, detecting the amplified ligation product, and identifying the target nucleotide sequence. Such coupling of the ligase detection reaction and the polymerase chain reaction permits multiplex detection of nucleic acid sequence difference.

主权项

1. A kit for identifying a plurality of polymorphic loci comprising:
a plurality of primary oligonucleotide primer sets, each primer set comprising (i) a first oligonucleotide primer comprising a target-specific portion and a 5′ upstream secondary primer-specific portion and (ii) a second oligonucleotide primer comprising a target-specific portion and a 5′ upstream, secondary primer-specific portion; a polymerase; and an oligonucleotide secondary primer set comprising (i) a first secondary primer containing the same sequence as the 5′ upstream primer-specific portion of the first primary oligonucleotide primer and (ii) a secondary primer containing the same sequence as the 5′ upstream portion of the second primary oligonucleotide primer, wherein the first or second secondary primers comprise a detectable label.