CHEMICAL COMPOSITIONS AND METHODS OF USE
The present invention relates to sequencing probes, methods, kits, and apparatuses that provide enzyme-free, amplification-free, and library-free nucleic acid sequencing that has long-read-lengths and with low error rate.
1 . A kit comprising a first plurality of oligonucleotides, wherein the oligonucleotides in the first plurality comprise at least one of the following chemical formulas:
2 . The kit of claim 1 , wherein the oligonucleotides in the first plurality comprise:
3 . The kit of claim 1 , wherein the oligonucleotides in the first plurality comprise:
4 . The kit of claim 1 , wherein the oligonucleotides in the first plurality comprise:
5 . The kit of claim 1 , wherein the oligonucleotides in the first plurality comprise:
6 . The kit of claim 1 , wherein the oligonucleotides in the first plurality comprise:
7 . The kit of claim 1 , wherein the oligonucleotides in the first plurality comprise:
8 . The kit of claim 1 , wherein the oligonucleotides in the first plurality comprise:
9 . The kit of claim 1 , wherein the oligonucleotides in the first plurality comprise:
10 . The kit of claim 1 , wherein the oligonucleotides in the first plurality comprise:
11 . The kit of claim 1 , wherein the kit further comprises at least a second plurality of oligonucleotides, wherein the oligonucleotides in the second plurality comprise a target binding domain and a barcode domain;
wherein said target binding domain comprises at least 12 nucleotides and is capable of binding a target nucleic acid;
wherein said barcode domain comprises a synthetic backbone, said barcode domain comprising at least four attachment positions, each attachment position comprising at least one attachment region, said attachment region comprising at least one nucleic acid sequence capable of being bound by a complementary nucleic acid molecule,
wherein the at least four attachment positions correspond to the sequence of the target binding domain and wherein each of the at least four attachment positions have a different nucleic acid sequence, and
wherein said nucleic acid sequences of each position of the at least four attachment positions determine the identity of the target nucleic acid that is bound by said target binding domain one nucleotide in said target nucleic acid that is bound by said target binding domain.
12 . A method of detecting at least one target nucleic acid in a sample comprising:
(1) contacting the sample with the second plurality of oligonucleotides of the kit of claim 11 such that at least one oligonucleotide from the second plurality binds to the at least one target nucleic acid;
(2) contacting the sample with the first plurality of oligonucleotides of the kit of claim 11 such that at least one oligonucleotide in the first plurality binds to the at least one oligonucleotide in the second plurality that is bound to the at least one target nucleic acid; and
(3) detecting the at least one oligonucleotide in the first plurality bound to the at least one oligonucleotide in the second plurality, thereby detecting the at least one target nucleic acid.