Chemical methods for producing tagged nucleotides
AssigneesThe Trustees of Columbia University in the City of New York, Genia Technologies, Inc.
InventorsCarl W. Fuller, Shiv Kumar, Jingyue Ju, Randall Davis, Roger Chen
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Abstract
This disclosure provides systems and methods for attaching nanopore-detectable tags to nucleotides. The disclosure also provides methods for sequencing nucleic acids using the disclosed tagged nucleotides.
Claims20
A tagged nucleotide, comprising a poly-phosphate moiety having a terminal phosphate at the 5′ position of the nucleotide, and a tag covalently coupled to the terminal phosphate of the nucleotide by a triazole, a 1,2-diazine, a disulfide, a secondary amine, a hydrazone, a thio-acetamide, or a maleimide-thioadduct,
- wherein the tag comprises an oligonucleotide, wherein:
- (i) the oligonucleotide comprises a non-naturally occurring internucleotide linkage;
- (ii) the 5′-end of the oligonucleotide is covalently coupled to the terminal phosphate and the 3′ terminus of the oligonucleotide has a chemical modification which protects it from exonuclease degradation, or the 3′-end of the oligonucleotide is covalently coupled to the terminal phosphate and the 5′ terminus of the oligonucleotide has a chemical modification which protects it from exonuclease degradation;
- or
- (iii) the oligonucleotide comprises at least one abasic spacer (dSp).
Dependencies are inferred from the claim text.