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US 10,907,149 B2Granted
Methods for targeted genomic analysis
AssigneeResolution Bioscience, Inc.
InventorsChristopher K. Raymond, Christopher D. Armour, Lee P. Lim
Life of this patent
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Abstract
The invention provides a method for genetic analysis in individuals that reveals both the genetic sequences and chromosomal copy number of targeted and specific genomic loci in a single assay. The present invention further provide methods for the sensitive and specific detection of target gene sequences and gene expression profiles.
Claims10
A method for determining copy number of a specific genomic target region comprising:
- (a) hybridizing a tagged genomic library with a multifunctional capture probe module to form a complex,
- (i) wherein the multifunctional capture probe module selectively hybridizes to a specific genomic target region in the genomic library and wherein the multifunctional capture probe module comprises a tail portion comprising a PCR binding site,
- (ii) wherein the tagged genomic library comprises genomic DNA ligated to a multifunctional adaptor module comprising a first region comprising a random nucleic acid tag sequence;
- a second region comprising a sample code sequence;
- and a third region comprising a tail portion comprising a PCR primer sequence, and
- (iii) wherein the sample code sequence identifies the sample;
- (b) isolating the tagged genomic library-multifunctional capture probe module complex from a);
- (c) performing 3′-5′ exonuclease enzymatic processing on the isolated tagged genomic library-multifunctional capture probe module complex from b) using an enzyme with 3′-5′ exonuclease activity to remove the single stranded 3′ ends;
- (d) performing a PCR reaction on the enzymatically processed complex from c) wherein a tail portion of the multifunctional capture probe module is replicated to generate a hybrid nucleic acid molecule, wherein the hybrid nucleic acid molecule comprises the genomic target region capable of hybridizing to the multifunctional capture probe module and a complement of the multifunctional capture probe module tail sequence;
- (e) performing PCR amplification of the hybrid nucleic acid molecule in d);
- and
- (f) quantitating the PCR reaction in e), wherein the quantitation comprises determination of copy number of the specific genomic target region.
9 more elements
A method for determining copy number of a specific genomic target region comprising:
- (a) hybridizing a tagged genomic library with a multifunctional capture probe module to form a complex,
- (i) wherein the multifunctional capture probe module selectively hybridizes to a specific genomic target region in the genomic library,
- (ii) wherein the tagged genomic library comprises genomic DNA ligated to a multifunctional adaptor module comprising a first region comprising a random nucleic acid tag sequence;
- a second region comprising a sample code sequence;
- and
- (iii) wherein the sample code sequence identifies the sample;
- (b) isolating the tagged genomic library-multifunctional capture probe hybrid module complex from a);
- (c) performing 5′ to 3′ DNA polymerase extension of the multifunctional capture probe on the complex from b) to replicate a region of the captured, tagged genomic target region that is 3′ of the multifunctional capture probe to generate a hybrid nucleic acid molecule, wherein the hybrid nucleic acid molecule comprises the multifunctional capture probe hybrid module and a complement of the region of the tagged genomic target region that is located in the 3′ direction from the location where the multifunctional capture probe hybrid module hybridizes to the genomic target region;
- and
- (d) performing PCR amplification of the hybrid nucleic acid molecule in c);
- and
- (e) quantitating the PCR reaction in d), wherein the quantitation comprises determination of copy number of the specific genomic target region.
9 more elements
Dependencies are inferred from the claim text.