Protocol for detecting cancer in blood using targeted DNA methylation
Here, we present a protocol for detecting and differentiating colorectal and gastric cancers using plasma cell-free DNA (cfDNA) with targeted methylated CpG tandem amplification and sequencing (tMCTA-seq), a targeted DNA methylation sequencing approach. We describe steps for sample collection, DNA extraction, bisulfite conversion, and targeting 110 methylated sites by coupling locus-specific primers with a universal CGCGCGG primer. We then detail an optional workflow that adapts tMCTA-seq to other cancer types. This approach achieves high sensitivity, below one haploid-genome equivalent, enabling non-invasive cancer detection.
Authors
- Xin Zhou (ORCID: https://orcid.org/0000-0002-7184-1101)
- Kun Qian (ORCID: https://orcid.org/0000-0003-0033-5903)
- Lu Wen (ORCID: https://orcid.org/0000-0002-1773-1876)
- Yongqu Lu (ORCID: https://orcid.org/0000-0001-9363-3189)
- Zhaoyu Jiang (ORCID: https://orcid.org/0000-0002-6865-7602)
- Yuqing Guo (ORCID: https://orcid.org/0009-0000-3155-7707)
- Xiaomeng Liu (ORCID: https://orcid.org/0000-0002-3288-212X)
- Jie Ren
- Wei Fu
- Renjie Luo
Institutions
- Peking University (CN)
- China-Japan Friendship Hospital (CN)
- Sichuan Cancer Hospital (CN)
- Ministry of Education (ET)
- Shandong First Medical University (CN)
Publication Details
- Journal
- STAR Protocols
- Published
- 2026-08-20
- DOI
- https://doi.org/10.1016/j.xpro.2026.104791
- Primary Topic
- Cancer Genomics and Diagnostics
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- Natural Science Foundation of Shandong Province