TY - CHAP
T1 - Post-bisulfite Adaptor Tagging Based on an ssDNA Ligation Technique (tPBAT)
AU - Miura, Fumihito
AU - Shibata, Yukiko
AU - Miura, Miki
AU - Ito, Takashi
N1 - Funding Information:
This work was supported by JSPS KAKENHI through grants to F.M. (20H03243) and T.I. (17H06305) and AMED Platform Project for Supporting Drug Discovery and Life Science Research (Basis for Supporting Innovative Drug Discovery and Life Science Research (BINDS)) through a grant to T.I. (Grant Number JP20am0101103).
Publisher Copyright:
© 2023, The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.
PY - 2023
Y1 - 2023
N2 - Post-bisulfite adaptor tagging (PBAT) is a concept that enables the preparation of an efficient sequencing library from bisulfite-treated DNA, and it also means the protocol implemented the concept. Although the previous PBAT or rPBAT was sensitive enough for single-cell methylome analysis, the protocol had several drawbacks owing to the repeated random priming reactions. To resolve these problems, we developed a unique single-strand DNA ligation technique, termed TACS ligation, and established a new protocol called tPBAT. With tPBAT, the data quality improved, with a longer insert and higher mapping rate than that obtained with rPBAT. In addition, paired-end sequencing and indexing were supported by the default. In this chapter, the tPBAT protocol is introduced, and a thorough description of its application to small samples is provided.
AB - Post-bisulfite adaptor tagging (PBAT) is a concept that enables the preparation of an efficient sequencing library from bisulfite-treated DNA, and it also means the protocol implemented the concept. Although the previous PBAT or rPBAT was sensitive enough for single-cell methylome analysis, the protocol had several drawbacks owing to the repeated random priming reactions. To resolve these problems, we developed a unique single-strand DNA ligation technique, termed TACS ligation, and established a new protocol called tPBAT. With tPBAT, the data quality improved, with a longer insert and higher mapping rate than that obtained with rPBAT. In addition, paired-end sequencing and indexing were supported by the default. In this chapter, the tPBAT protocol is introduced, and a thorough description of its application to small samples is provided.
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U2 - 10.1007/978-1-0716-2724-2_2
DO - 10.1007/978-1-0716-2724-2_2
M3 - Chapter
C2 - 36173563
AN - SCOPUS:85139379199
T3 - Methods in Molecular Biology
SP - 21
EP - 37
BT - Methods in Molecular Biology
PB - Humana Press Inc.
ER -