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.
CITATION STYLE
Miura, F., Shibata, Y., Miura, M., & Ito, T. (2023). Post-bisulfite Adaptor Tagging Based on an ssDNA Ligation Technique (tPBAT). In Methods in Molecular Biology (Vol. 2577, pp. 21–37). Humana Press Inc. https://doi.org/10.1007/978-1-0716-2724-2_2
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