Triazole linking for preparation of a next-generation sequencing library from single-stranded DNA

Nucleic Acids Res. 2018 Sep 19;46(16):e95. doi: 10.1093/nar/gky452.


Next-generation sequencing of single-stranded DNA (ssDNA) is attracting increased attention from a wide variety of research fields. Accordingly, various methods are actively being tested for the efficient adaptor-tagging of ssDNA. We conceived a novel chemo-enzymatic method termed terminal deoxynucleotidyl transferase (TdT)-assisted, copper-catalyzed azide-alkyne cycloaddition (CuAAC)-mediated ssDNA ligation (TCS ligation). In this method, TdT is used to incorporate a single 3'-azide-modified dideoxyribonucleotide onto the 3'-end of target ssDNA, followed by CuAAC-mediated click ligation of the azide-incorporated 3'-end to a 5'-ethynylated synthetic adaptor. This report presents the first proof-of-principle application of TCS ligation with its use in the preparation of a next-generation sequencing library.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alkynes / chemistry
  • Azides / chemistry
  • Click Chemistry
  • Copper / chemistry
  • Cycloaddition Reaction
  • DNA Nucleotidylexotransferase / chemistry
  • DNA Nucleotidylexotransferase / metabolism
  • DNA, Single-Stranded*
  • Gene Library*
  • High-Throughput Nucleotide Sequencing / methods*
  • Micrococcal Nuclease / genetics
  • Micrococcal Nuclease / metabolism
  • Oligodeoxyribonucleotides / genetics
  • Oligodeoxyribonucleotides / metabolism
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / metabolism
  • Sequence Analysis, DNA / methods
  • Triazoles / chemistry*


  • Alkynes
  • Azides
  • DNA, Single-Stranded
  • Oligodeoxyribonucleotides
  • Triazoles
  • Copper
  • DNA Nucleotidylexotransferase
  • Micrococcal Nuclease