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.
| Original language | English |
|---|---|
| Pages (from-to) | e95 |
| Journal | Nucleic acids research |
| Volume | 46 |
| Issue number | 16 |
| DOIs | |
| Publication status | Published - Sep 19 2018 |
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All Science Journal Classification (ASJC) codes
- Genetics
Cite this
Triazole linking for preparation of a next-generation sequencing library from single-stranded DNA. / Miura, Fumihito; Fujino, Tomoko; Kogashi, Kanako; Shibata, Yukiko; Miura, Miki; Isobe, Hiroyuki; Ito, Takashi.
In: Nucleic acids research, Vol. 46, No. 16, 19.09.2018, p. e95.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Triazole linking for preparation of a next-generation sequencing library from single-stranded DNA
AU - Miura, Fumihito
AU - Fujino, Tomoko
AU - Kogashi, Kanako
AU - Shibata, Yukiko
AU - Miura, Miki
AU - Isobe, Hiroyuki
AU - Ito, Takashi
PY - 2018/9/19
Y1 - 2018/9/19
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85060116179&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85060116179&partnerID=8YFLogxK
U2 - 10.1093/nar/gky452
DO - 10.1093/nar/gky452
M3 - Article
C2 - 29846671
AN - SCOPUS:85060116179
VL - 46
SP - e95
JO - Nucleic Acids Research
JF - Nucleic Acids Research
SN - 0305-1048
IS - 16
ER -