Methods and materials for assessing allelic imbalance
Methods and systems for detecting allelic imbalance using nucleic acid sequencing are provided.
1. An in vitro method for detecting homozygosity at a plurality of single nucleotide polymorphism loci, comprising:
(1) providing a formalin-fixed paraffin-embedded sample comprising at least one tumor cell obtained from a patient;
(2) enriching the sample in (1) for test DNA molecules each comprising at least one locus from the plurality of single nucleotide polymorphism loci, wherein the plurality of single nucleotide polymorphism loci comprises at least 5,000 single nucleotide polymorphism loci and wherein there is at least one single nucleotide polymorphism locus located on average every 500 kb within each chromosome; and
(3) sequencing the test DNA molecules to determine whether each locus in the plurality of single nucleotide polymorphism loci is homozygous.
2. The method of claim 1 , wherein the plurality of single nucleotide polymorphism loci comprises at least 10,000 single nucleotide polymorphism loci.
3. The method of claim 1 , wherein the plurality of single nucleotide polymorphism loci comprises at least 50,000 single nucleotide polymorphism loci.
4. The method of claim 1 , wherein there is at least one single nucleotide polymorphism locus located on average every 100 kb within each chromosome.
5. The method of claim 1 , wherein there is at least one single nucleotide polymorphism locus located on average every 50 kb within each chromosome.
6. The method of claim 1 , wherein there is at least one single nucleotide polymorphism locus located on average every 10 kb within each chromosome.
7. The method of claim 1 , wherein the genomic spacing of the plurality of single nucleotide polymorphism loci is less than or equal to 50%.
8. The method of claim 1 , wherein the genomic spacing of the plurality of single nucleotide polymorphism loci is less than or equal to 25%.
9. The method of claim 1 , wherein the genomic spacing of the plurality of single nucleotide polymorphism loci is less than or equal to 10%.
10. A system for detecting homozygosity at a plurality of single nucleotide polymorphism loci, comprising:
(1) a sample analyzer for
(a) enriching a formalin-fixed paraffin-embedded sample comprising at least one tumor cell obtained from a patient for test DNA molecules each comprising at least one locus from the plurality of single nucleotide polymorphism loci, wherein the plurality of single nucleotide polymorphism loci comprises at least 5,000 single nucleotide polymorphism loci and wherein there is at least one single nucleotide polymorphism locus located on average every 500 kb within each chromosome; and
(b) sequencing the test DNA molecules to produce a plurality of quantitative signals representing the alleles for each locus in the plurality of single nucleotide polymorphism loci present in the test DNA molecules; and
(2) a computer program for analyzing the plurality of quantitative signals to determine whether each locus in the plurality of single nucleotide polymorphism loci is homozygous.
11. The method of system 10 , wherein the plurality of single nucleotide polymorphism loci comprises at least 10,000 single nucleotide polymorphism loci.
12. The method of system 10 , wherein the plurality of single nucleotide polymorphism loci comprises at least 50,000 single nucleotide polymorphism loci.
13. The method of system 10 , wherein there is at least one single nucleotide polymorphism locus located on average every 100 kb within each chromosome.
14. The method of system 10 , wherein there is at least one single nucleotide polymorphism locus located on average every 50 kb within each chromosome.
15. The method of system 10 , wherein there is at least one single nucleotide polymorphism locus located on average every 10 kb within each chromosome.
16. The method of system 10 , wherein the genomic spacing of the plurality of single nucleotide polymorphism loci is less than or equal to 50%.
17. The method of system 10 , wherein the genomic spacing of the plurality of single nucleotide polymorphism loci is less than or equal to 25%.
18. The method of system 10 , wherein the genomic spacing of the plurality of single nucleotide polymorphism loci is less than or equal to 10%.