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A capture methyl-seq protocol with improved efficiency and cost-effectiveness using pre-pooling and enzymatic conversion

Research Square
2022
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Article Description

Objective The opportunities for sequencing-based methylome analysis of clinical samples are increasing. To reduce its cost and the amount of genomic DNA required for library preparation, we aimed to establish a capture methyl-seq protocol, which adopts pre-pooling of multiple libraries before hybridization capture and TET2/APOBEC-mediated conversion of unmethylated cytosine to thymine. Results We compared a publicly available dataset generated by the standard protocol of SureSelect XT Human Methyl-Seq Kit and our dataset obtained by its modified protocol that adopted sample pre-pooling and enzymatic conversion. We confirmed that the quality of DNA methylation data was comparable between the two datasets. As our protocol, EMCap, is more cost-effective and reduces the amount of input genomic DNA, it would serve as a better choice for clinical methylome sequencing.

Bibliographic Details

Keita Hasegawa; Kazuhiko Nakabayashi; Keisuke Ishiwata; Kenichiro Hata; Yoshifumi Kasuga; Mamoru Tanaka

Research Square Platform LLC

Biochemistry, Genetics and Molecular Biology; Immunology and Microbiology; Medicine; Neuroscience; Psychology; Dentistry

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