异常的 DNA 甲基化是癌症的普遍特征。在这里，我们提出了一种从各种人类癌症生物样本（包括永生化细胞系、新鲜冷冻手术切除和福尔马林固定石蜡包埋组织）生成高质量基因组规模 DNA 甲基化测序数据的协议。我们描述了 DNA 提取注意事项、简化代表性亚硫酸氢盐测序、数据处理和质量控制以及下游数据分析和集成的步骤。该协议也适用于其他人类疾病和其他生物体的甲基组分析。有关该协议的使用和执行的完整详细信息，请参阅 Rodger 等人。 (2023).1.版权所有 © 2023 作者。由爱思唯尔公司出版。保留所有权利。

Aberrant DNA methylation is a universal feature of cancer. Here, we present a protocol for generating high-quality genome-scale DNA methylation sequencing data from a variety of human cancer biospecimens including immortalized cell lines, fresh-frozen surgical resections, and formalin-fixed paraffin-embedded tissues. We describe steps for DNA extraction considerations, reduced representation bisulfite sequencing, data processing and quality control, and downstream data analysis and integration. This protocol is also applicable for other human diseases and methylome profiling in other organisms. For complete details on the use and execution of this protocol, please refer to Rodger et al. (2023).1.Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.