A capture methyl-seq protocol with improved efficiency and cost-effectiveness using pre-pooling and enzymatic conversion
説明
<jats:title>Abstract</jats:title><jats:sec> <jats:title>Objective</jats:title> <jats:p>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.</jats:p> </jats:sec><jats:sec> <jats:title>Results</jats:title> <jats:p>We compared a publicly available dataset generated by the standard Agilent protocol of SureSelect XT Human Methyl-Seq Kit and our dataset obtained by our modified protocol, EMCap, 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.</jats:p> </jats:sec>
収録刊行物
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- BMC Research Notes
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BMC Research Notes 16 (1), 2023-07-06
Springer Science and Business Media LLC
