Comparative analysis of improved m6A sequencing based on antibody optimization for low-input samples

作者：Jiafeng Lu, Wenjuan Xia, Jincheng Li, Liya Zhang, Chunfeng Qian, Hong Li, Boxian Huang · 发表于：Scientific Reports · 年份：2025 · DOI：10.1038/s41598-025-85150-8 · 被引用次数：6 · 研究领域：RNA modifications and cancer、Cancer-related gene regulation、HVDC Systems and Fault Protection

The most effective method for mapping N6-methyladenosine (m 6 A) is m 6 A RNA immunoprecipitation sequencing (MeRIP-seq). The quality of MeRIP-seq relies on various factors, with the anti-m 6 A antibody being a crucial determinant. However, comprehensive research on anti-m 6 A antibody selection and optimal concentrations for different tissues has been limited. In this study, we optimized the concentration of five different anti-m 6 A antibodies across various tissues. Our findings demonstrated that 5 µg of Millipore antibodies (ABE572 and MABE1006) performed well, starting from 15 µg total RNA from the liver, while 1.25 µg of Cell Signaling Technology antibodies (CST) (#56593) was suitable for low-input total RNA. In summary, we provide a significant guideline for anti-m 6 A antibody selection in MeRIP sequencing for different tissues, especially in the context of low-input RNA.