Multiplexed sequential ligation-based detection of genetic variants

摘要

The present invention provides multiplexed sequential ligation-based analysis of genetic variants in a mixed sample, including copy number variations and single nucleotide polymorphisms. The invention employs the techniques of sequential ligation and amplification.

主权项

1. A method for identifying a genomic region of interest from a single source in a sample comprising DNA from two different sources, comprising the steps of:
providing a sample comprising DNA from two different sources; introducing to the sample a first set of oligonucleotide probes comprising a first fixed sequence oligonucleotide complementary to a 3′ region in a genomic region of interest and a second fixed sequence oligonucleotide complementary to a 5′ region in the genomic region of interest; hybridizing the first set of oligonucleotide probes to the genomic region of interest in the sample; ligating the hybridized oligonucleotides of the first set of oligonucleotide probes to create first ligation products complementary to the genomic region of interest; introducing to the first ligation products a second set of oligonucleotide probes comprising a first fixed sequence oligonucleotide complementary to a 3′ region in the first ligation product and a second fixed sequence oligonucleotide complementary to a 5′ region in the first ligation product; hybridizing the second set of oligonucleotide probes to the first ligation products; ligating the hybridized oligonucleotides of the second set of oligonucleotide probes to create second ligation products complementary to the first ligation products, wherein junctions for ligation for the second set of oligonucleotides probes are different than junctions for ligation for the first set of oligonucleotides probes; amplifying the second ligation products to create amplification products; and analyzing the amplification products, wherein the analysis of the amplification products identifies the genomic region of interest from the single source in the sample.