CHEMICAL COMPOSITIONS AND METHODS OF USING SAME
The present disclosure relates to chemical compositions, kits, and apparatuses and methods for using these compositions, kits and apparatuses in various assays.
1 . (canceled)
2 . A probe comprising:
a target binding domain and a barcode domain,
wherein the barcode domain comprises a synthetic backbone, the barcode domain including at least two attachment positions, each attachment position including at least one nucleic acid sequence that hybridizes to a complementary nucleic acid molecule, and
wherein the synthetic backbone includes L-DNA,
wherein each nucleotide of the at least one nucleic acid sequence of each attachment region is L-DNA, and
wherein the at least two attachment positions correspond to a sequence of the target binding domain; and
at least one cleavable linker positioned between the target binding domain and the barcode domain.
3 . The probe of claim 2 , wherein each nucleotide of the target binding domain is D-DNA.
4 . The probe of claim 2 , wherein the target binding domain comprises a sequence corresponding to a known RNA sequence.
5 . The probe of claim 2 , wherein the target binding domain comprises a sequence corresponding to a known DNA sequence.
6 . The probe of claim 2 , wherein the target binding domain is at least 12 nucleotides in length.
7 . The probe of claim 2 , wherein the barcode domain comprises at least three attachment positions.
8 . The probe of claim 2 , wherein the barcode domain comprises at least four attachment positions.
9 . The probe of claim 2 , further comprising:
a reporter probe including a reporter probe sequence corresponding to the nucleic acid sequence of a first of the at least one attachment positions, the reporter probe including at least one fluorescent molecule.
10 . The probe of claim 2 , further comprising:
a first reporter probe including a sequence corresponding to the nucleic acid sequence of a first of the at least one attachment positions, the first reporter probe including at least one first fluorescent molecule; and
a second reporter probe including a sequence corresponding to the nucleic acid sequence of a second of the at least one attachment positions, the second reporter probe including at least one second fluorescent molecule.
11 . The probe of claim 10 , wherein the first fluorescent molecule and the second fluorescent molecule are spectrally resolvable.
12 . The probe of claim 2 , wherein the cleavable linker is a photocleavable linker.
13 . A method for identifying a target nucleic acid in a sample, comprising:
hybridizing the target binding domain of the probe of claim 2 to a target nucleic acid;
hybridizing a first reporter probe to at least one of the at least two attachment positions of the barcode domain, the first reporter probe including at least one fluorescent molecule;
identifying an emission spectrum from the at least one fluorescent molecule;
exposing the sample to light sufficient to break the cleavable linker, thereby removing at least one of the at least two attachment positions from the probe molecule; and
determining a location of the target nucleic acid based on the identified emission spectrum of the at least one fluorescent molecule.