This study reports high-resolution DNA methylation profiles of human chromosomes 6, 20, and 22 using bisulfite DNA sequencing. The analysis covers 12 different tissues and 1.9 million CpG sites, providing a comprehensive resource for understanding epigenetic regulation. Key findings include: 1. **High-resolution Methylation Profiles**: The study generated detailed methylation profiles for human chromosomes 6, 20, and 22, covering 1.9 million CpG sites from 12 different tissues. 2. **Evolutionary Conservation and Differentially Methylated Regions (DMRs)**: The analysis identified evolutionary conserved regions as the predominant sites for differential DNA methylation. Tissue-specific differentially methylated regions (T-DMRs) were found in 22% of amplicons, particularly in 5′-untranslated regions (5′-UTRs) and evolutionary conserved regions (ECRs). 3. **Promoter Methylation and Transcriptional Regulation**: Promoter methylation was analyzed using 'promoter-proxy' regions, including 5′-UTRs, transcription start sites (TSSs), and Sp1 binding sites. The study found that the core region surrounding the TSS is informative for promoter methylation, and Sp1 binding was associated with unmethylated and TSS-associated sites. 4. **Age and Sex-Dependent DNA Methylation**: The study controlled for age and sex, finding minimal effects on methylation levels. However, significant differences were observed between different cell types, such as CD4+ and CD8+ lymphocytes. 5. **Conservation of DNA Methylation**: The methylation profiles of orthologous amplicons between human and mouse tissues showed high conservation, with 69.4% of amplicons differing by less than 20% methylation. 6. **Implications for Epigenetic Regulation**: The findings suggest that DNA methylation is more stable than previously thought and play a crucial role in tissue-specific transcription and epigenetic plasticity. This study contributes to the understanding of the human epigenome and provides a valuable resource for further research on epigenetic regulation and disease.This study reports high-resolution DNA methylation profiles of human chromosomes 6, 20, and 22 using bisulfite DNA sequencing. The analysis covers 12 different tissues and 1.9 million CpG sites, providing a comprehensive resource for understanding epigenetic regulation. Key findings include: 1. **High-resolution Methylation Profiles**: The study generated detailed methylation profiles for human chromosomes 6, 20, and 22, covering 1.9 million CpG sites from 12 different tissues. 2. **Evolutionary Conservation and Differentially Methylated Regions (DMRs)**: The analysis identified evolutionary conserved regions as the predominant sites for differential DNA methylation. Tissue-specific differentially methylated regions (T-DMRs) were found in 22% of amplicons, particularly in 5′-untranslated regions (5′-UTRs) and evolutionary conserved regions (ECRs). 3. **Promoter Methylation and Transcriptional Regulation**: Promoter methylation was analyzed using 'promoter-proxy' regions, including 5′-UTRs, transcription start sites (TSSs), and Sp1 binding sites. The study found that the core region surrounding the TSS is informative for promoter methylation, and Sp1 binding was associated with unmethylated and TSS-associated sites. 4. **Age and Sex-Dependent DNA Methylation**: The study controlled for age and sex, finding minimal effects on methylation levels. However, significant differences were observed between different cell types, such as CD4+ and CD8+ lymphocytes. 5. **Conservation of DNA Methylation**: The methylation profiles of orthologous amplicons between human and mouse tissues showed high conservation, with 69.4% of amplicons differing by less than 20% methylation. 6. **Implications for Epigenetic Regulation**: The findings suggest that DNA methylation is more stable than previously thought and play a crucial role in tissue-specific transcription and epigenetic plasticity. This study contributes to the understanding of the human epigenome and provides a valuable resource for further research on epigenetic regulation and disease.