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

Nucleic Acids Research
Fumihito MiuraTakashi Ito

Abstract

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.

References

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Citations

Sep 4, 2019·Chemistry, an Asian Journal·Tomoko FujinoHiroyuki Isobe
Dec 25, 2019·Bioconjugate Chemistry·Michael KollaschinskiSascha Serdjukow
Sep 22, 2021·Journal of the American Chemical Society·Sven EppleTom Brown

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Datasets Mentioned

BETA
GSE110666
SRP133409

Methods Mentioned

BETA
PCR
DNA-seq
MNase-seq
electrophoresis
Illumina sequencing
footprinting
RNA-seq

Software Mentioned

ClickSeq
bowtie
in
house
SPRI

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