DNA 包含存储于遗传和表观遗传基础中的分子信息，这两者对于我们理解生物学至关重要。大多数 DNA测序方法要么研究遗传学，要么关注表观遗传学，因此无法捕捉完整的信息。广泛用于检测表观遗传基础的方法无法捕捉常见的 C-to-T突变，也无法区分 5-甲基胞嘧啶和 5-羟甲基胞嘧啶。我们提出了一种单碱基分辨率测序方法，可以在一次工作流程中同时测序完整的遗传信息和最常见的两种胞嘧啶修饰。复制 DNA 后，碱基经过酶促转化，将原始链和副本链的碱基进行耦合解码，提供分阶段的数码输出。我们对人类基因组 DNA 和来自癌症患者血样的细胞游离 DNA 进行了演示。这种方法准确、需要低 DNA输入量，并具有简单的工作流程和分析管道。同时、分阶段阅读遗传和表观遗传基础可以提供关于存储于基因组中的信息更完整的图像，并在生物医学领域得到广泛应用。 © 2023. 作者（或合著者)。

DNA comprises molecular information stored in genetic and epigenetic bases, both of which are vital to our understanding of biology. Most DNA sequencing approaches address either genetics or epigenetics and thus capture incomplete information. Methods widely used to detect epigenetic DNA bases fail to capture common C-to-T mutations or distinguish 5-methylcytosine from 5-hydroxymethylcytosine. We present a single base-resolution sequencing methodology that sequences complete genetics and the two most common cytosine modifications in a single workflow. DNA is copied and bases are enzymatically converted. Coupled decoding of bases across the original and copy strand provides a phased digital readout. Methods are demonstrated on human genomic DNA and cell-free DNA from a blood sample of a patient with cancer. The approach is accurate, requires low DNA input and has a simple workflow and analysis pipeline. Simultaneous, phased reading of genetic and epigenetic bases provides a more complete picture of the information stored in genomes and has applications throughout biomedicine.© 2023. The Author(s).