Department of Maternal-Fetal Biology, National Center for Child Health and Development, 2-10-1 Okura, Setagaya-ku, Tokyo, 157-8535, Japan.
Department of Obstetrics and Gynecology, Keio University School of Medicine, Shinjuku, Tokyo, 160-0016, Japan.
BMC Res Notes. 2023 Jul 6;16(1):141. doi: 10.1186/s13104-023-06401-3.
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.
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.
基于测序的甲基组分析在临床样本中的应用机会正在增加。为了降低成本和文库制备所需的基因组 DNA 量，我们旨在建立一种捕获甲基化测序方案，该方案采用杂交捕获前的多个文库预池化和 TET2/APOBEC 介导的未甲基化胞嘧啶向胸腺嘧啶的转化。
我们比较了一个公开的数据集，该数据集是由标准的 Agilent 方案的 SureSelect XT Human Methyl-Seq Kit 生成的，和我们通过我们的修改的方案 EMCap 获得的数据集，该方案采用了样品预池化和酶转化。我们证实了两个数据集的 DNA 甲基化数据质量是可比的。由于我们的方案 EMCap 更具成本效益，并且减少了输入基因组 DNA 的量，因此它将成为临床甲基组测序的更好选择。
