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Chemical methods for producing tagged nucleotides

  • US 10,240,195 B2
  • Filed: 03/23/2015
  • Issued: 03/26/2019
  • Est. Priority Date: 03/24/2014
  • Status: Active Grant
First Claim
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1. 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).

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