Methods for detecting and identifying genomic nucleic acids
The present invention relates to a method of identifying a target genomic nucleic acid sequence including hybridizing a set of probes to the target genomic nucleic acid sequence, wherein the set of probes has a unique associated barcode sequence for identification of the target genomic nucleic acid sequence, wherein each probe of the set includes (1) a complementary sequence complementary to a first strand of the target genomic nucleic acid sequence and (2) the associated barcode sequence or a portion of the associated barcode sequence, sequencing the associated barcode sequence from probes hybridized to the target genomic nucleic acid sequence using a fluorescence-based sequencing method, and identifying the target genomic nucleic acid sequence by the sequenced barcode sequence.
1. A method of identifying a target genomic nucleic acid sequence comprising
hybridizing a set of probes to the target genomic nucleic acid sequence, wherein the set of probes has a unique associated barcode sequence for identification of the target genomic nucleic acid sequence, wherein each probe of the set includes (1) a complementary sequence complementary to a first strand or a second strand of the target genomic nucleic acid sequence and (2) the associated barcode sequence or a portion of the associated barcode sequence,
sequencing the associated barcode sequence from probes hybridized to the target genomic nucleic acid sequence using a fluorescence-based sequencing method, and
identifying the target genomic nucleic acid sequence by the sequenced barcode sequence.
2. The method of claim 1 wherein the fluorescence-based sequencing method is sequencing by synthesis and the probe further includes a priming sequence for annealing a sequencing primer, wherein the sequencing primer is extended by a DNA polymerase using reversible terminator fluorescently encoded dNTPs to generate a fluorescent signal corresponding to the barcode.
3. The method of claim 1 wherein the fluorescence-based sequencing method is sequencing by ligation and the probe further includes a priming sequence for annealing a sequencing primer, wherein the sequencing primer is extended in either the 5′ or 3′ direction by a DNA ligase using fluorescently encoded oligonucleotides to generate a fluorescent signal corresponding to the barcode.
4. The method of claim 1 wherein the fluorescence-based sequencing method is sequencing by hybridization and the probe further includes a first nucleic acid sequence complementary to a labeled oligonucleotide which hybridizes to the first nucleic acid sequence.
5. The method of claim 1 wherein the fluorescence-based sequencing method is sequencing by cyclic reversible polymerization hybridization chain reaction.
6. The method of claim 1 wherein a plurality of probes having a portion of the associated barcode sequence constitute a complete associated barcode sequence.
7. The method of claim 1 wherein the complementary sequence has a nucleotide length of between 5 and 10,000 bases.
8. The method of claim 1 wherein the complementary sequence has a nucleotide length of between 15 and 1,000 bases.
9. The method of claim 1 wherein the complementary sequence has a nucleotide length of between 20 and 80 bases.
10. The method of claim 1 wherein the probe includes one or more spacer sequences separating the complementary sequence from the associated barcode sequence or portion of the associate barcode sequence.
11. The method of claim 1 wherein the probe includes one or more spacer sequences comprising a plurality of dT nucleotides separating the complementary sequence from the associated barcode sequence or portion of the associate barcode sequence.
12. The method of claim 1 wherein the probe includes one or more spacer sequences separating functional sequences of the probe including the complementary sequence, the associated barcode sequence or portion of the associate barcode sequence, and a priming sequence.
13. The method of claim 1 wherein the probe includes one or more spacer sequences comprising a plurality of dT nucleotides separating functional sequences of the probe including the complementary sequence, the associated barcode sequence or portion of the associate barcode sequence, and a priming sequence.
14. The method of claim 1 wherein the target genomic nucleic acid has a length of between 10 bp and 1,000,000,000 bp.
15. The method of claim 1 wherein the target genomic nucleic acid has a length of between 20 bp and 1,000,000 bp.
16. The method of claim 1 wherein the target genomic nucleic acid has a length of between 100 bp and 1,000,000 bp.
17. The method of claim 1 wherein the target genomic nucleic acid has a length of between 100,000 bp and 1,000,000,000 bp.
18. The method of claim 1 wherein the target genomic nucleic acid is a whole genome.
19. The method of claim 1 wherein the probe includes one or more additional barcode sequences for barcoding characteristics of the target genomic nucleic acid.
20. The method of claim 1 wherein the probes are oligopaints.
21. The method of claim 1 wherein the probes further include an attachment moiety for attachment to a matrix.
22. A method of multiplexing the identification of a plurality of target genomic nucleic acid sequences within genomic DNA comprising
hybridizing the genomic DNA with a plurality of probe sets corresponding to the plurality of target genomic nucleic acid sequences, wherein each probe set has a unique associated barcode sequence for identification of the corresponding target genomic nucleic acid sequence, wherein each probe of each probe set includes (1) a complementary sequence complementary to a first strand of the corresponding target genomic nucleic acid sequence and (2) the associated barcode sequence or a portion of the associated barcode sequence,
sequencing the associated barcode sequence from probes hybridized to the plurality of target genomic nucleic acid sequences using a fluorescence-based sequencing method, and
identifying the target genomic nucleic acid sequence by the sequenced barcode sequence.
23. The method of claim 22 wherein the plurality of target genomic nucleic acid sequences within genomic DNA is between 5 and 100,000,000 target genomic nucleic acid sequences.