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,
(2) the unique associated barcode sequence or a portion of the associated barcode sequence, and
(3) one or more additional barcode sequences;
sequencing the unique associated barcode sequence from probes hybridized to the target genomic nucleic acid sequence using a fluorescence-based sequencing method,
identifying the target genomic nucleic acid sequence by the sequenced unique barcode sequence.
2. The method of claim 1 , comprising detecting the one or more additional barcode sequences.
3. The method of claim 2 , wherein detecting the one or more additional barcode sequences comprises hybridizing a detectably labeled nucleic acid molecule to the one or more additional barcode sequences.
4. The method of claim 1 , wherein the probe comprises at least two different priming sequences for annealing at least two different sequencing primers.
5. The method of claim 4 , wherein at least one priming sequence is positioned to initiate sequencing of the unique associated barcode sequence, and wherein at least one different priming sequence is positioned to initiate sequencing of the one or more additional barcode sequences.
6. The method of claim 1 , wherein the probe comprises at least one priming sequence for annealing at least one amplification primer.
7. The method of claim 1 , wherein sequencing the unique associated barcode sequence comprises amplifying the probe, or a portion of the probe comprising the unique associated barcode sequence.
8. The method of claim 1 , wherein identifying the target genomic nucleic acid sequence comprises super-resolution microscopy.
9. The method of claim 1 , wherein identifying the target genomic nucleic acid sequence comprises Gaussian fitting of the fluorescence signal from the barcode.
10. The method of claim 1 , wherein after the hybridizing step, the method comprises contacting the target nucleic acid with an expandable matrix (ExM) gel.
11. 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 or a second strand of the target genomic nucleic acid sequence,
(2) the unique associated barcode sequence or a portion of the associated barcode sequence, and
(3) one or more additional barcode sequences;
sequencing the unique associated barcode sequence from probes hybridized to the plurality of target genomic nucleic acid sequence using a fluorescence-based sequencing method,
identifying the target genomic nucleic acid sequence by the sequenced unique barcode sequence.
12. The method of claim 11 , comprising detecting the one or more additional barcode sequences.
13. The method of claim 12 , wherein detecting the one or more additional barcode sequences comprises hybridizing a detectably labeled nucleic acid molecule to the one or more additional barcode sequences.
14. The method of claim 11 , wherein each probe in each set of probes comprises at least two different priming sequences for annealing at least two different sequencing primers.
15. The method of claim 14 , wherein at least one priming sequence is positioned to initiate sequencing of the unique associated barcode sequence, and wherein at least one different priming sequence is positioned to initiate sequencing of the one or more additional barcode sequences.
16. The method of claim 11 , wherein each probe in each set of probes comprises at least one priming sequence for annealing at least one amplification primer.
17. The method of claim 11 , wherein sequencing the unique associated barcode sequence comprises amplifying the probes, or a portion of the probes comprising the unique associated barcode sequence.
18. The method of claim 11 , wherein identifying the target genomic nucleic acid sequence comprises super-resolution microscopy.
19. The method of claim 11 , wherein identifying the target genomic nucleic acid sequence comprises Gaussian fitting of the fluorescence signal from the barcode.
20. The method of claim 11 , wherein after the hybridizing step, the method comprises contacting the target nucleic acid with an expandable matrix (ExM) gel.