AI Article Synopsis
- Next-generation sequencing of single-stranded DNA (ssDNA) is gaining popularity across various research areas, prompting the development of efficient methods for adaptor-tagging ssDNA.
- A new technique called TCS ligation combines terminal deoxynucleotidyl transferase (TdT) with copper-catalyzed azide-alkyne cycloaddition (CuAAC) to effectively tag ssDNA.
- The study demonstrates the successful application of TCS ligation in creating a sequencing library, marking it as a promising approach for future sequencing technologies.
Article 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.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6144795 | PMC |
| http://dx.doi.org/10.1093/nar/gky452 | DOI Listing |
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