5-Hydroxymethylcytosine (5hmC), known as the "sixth base" of the genome, is a critical epigenetic marker essential for understanding gene regulation, development, and disease mechanisms. CD BioSciences delivers precise and comprehensive 5hmC analysis services, enabling genome-wide mapping and locus-specific quantification to support your research in neurobiology, cancer, and developmental epigenetics.

Overview of 5-Hydroxymethylcytosine (5hmC)

5-Hydroxymethylcytosine (5hmC), often termed the "sixth base" of the genome, is a stable and functionally distinct oxidative derivative of 5-methylcytosine (5mC), generated primarily through the activity of ten-eleven translocation (TET) enzymes. Unlike 5mC—which is broadly associated with transcriptional repression—5hmC is enriched in actively transcribed gene bodies, enhancers, and poised regulatory regions, where it contributes to fine-tuning gene expression, facilitating active DNA demethylation, and regulating cellular differentiation. It is particularly abundant in neurons and embryonic stem cells, establishing it as a critical epigenetic mark in neurodevelopment and early embryogenesis. Due to its tissue-specific distribution and dynamic regulation, alterations in 5hmC levels are closely linked to neurodegenerative diseases, cognitive aging, and various cancers, making it a promising target for biomarker discovery and mechanistic studies in epigenetics.

Fig.1 Cytosine methylation cycle. (Ecsedi S, et al., 2018)

Our Services

As a leading provider of epigenetic analysis services, CD BioSciences offers end-to-end 5-hydroxymethylcytosine (5hmC) analysis solutions designed for accurate and biologically meaningful analysis. We combine selective chemistry and enzymatic methods with advanced sequencing technologies to accurately distinguish 5hmC, 5mC, and other cytosine modifications. Our services support a wide range of applications, from exploratory whole-genome analysis to candidate region validation, helping you gain deeper insights into 5hmC-mediated regulatory mechanisms.

5hmC Analysis Solutions by CD BioSciences

Our 5hmC analysis platform integrates multiple established and emerging technologies to ensure specificity, sensitivity, and reproducibility:

1. Global DNA Modification Quantification by LC-MS/MS
2. Global DNA Modification Quantification by ELISA
3. Tet-assisted Bisulfite Sequencing (TAB-seq) Service

1. Oxidative Bisulfite Sequencing (oxBS-seq) Service
2. hMeDIP-seq Service
3. Nanopore Sequencing Service
4. Other Technologies

Workflow of DNA 5hmC Analysis Service

Employing a seamless and stage-gated workflow, CD BioSciences' 5hmC analysis service is designed to ensure precision and reproducibility. From initial consultation to final reporting, each step is optimized to accurately capture and interpret this dynamic epigenetic mark, delivering reliable, publication-quality data.

Project Consultation & Design

We begin by collaborating with you to define your research objectives, select the most appropriate method (e.g., oxBS-Seq, TAB-Seq, or hMeDIP-Seq), and establish a clear project plan tailored to your sample type and biological questions.

Sample Preparation & QC

Your DNA samples undergo rigorous extraction and quality assessment, including quantification, purity checks, and integrity verification, to ensure they meet the stringent requirements for sensitive 5hmC detection.

Selective 5hmC Processing

This critical step applies method-specific chemistry or enzymology, such as oxBS-Seq or TAB-seq, to selectively distinguish 5hmC from 5mC, forming the foundation for accurate downstream analysis.

Library Preparation & Sequencing

Processed DNA is converted into sequencing libraries using optimized protocols, followed by high-depth sequencing on Illumina platforms to generate comprehensive and high-quality raw data.

Bioinformatics & Data Analysis

Our dedicated bioinformatics team processes sequencing data using specialized pipelines for precise 5hmC calling, differential analysis, genomic annotation, and functional enrichment, transforming raw data into interpretable biological insights.

Our Advantages

1. Specificity & Accuracy: We employ orthogonal methods (oxBS-seq, TAB-seq, chemical enrichment) to ensure precise discrimination between 5hmC, 5mC, and other modifications.
2. Sensitivity for Low-Abundance Marks: Optimized protocols enable detection of 5hmC even in tissues with low global levels.
3. End-to-End Expertise: From method selection to functional interpretation, our epigenetics specialists guide you throughout your project.
4. Broad Applicability: Services are validated for diverse samples, including brain tissue, blood, FFPE, and stem cells.
5. Integrated Multi-Omics Support: We offer correlative analysis with gene expression, chromatin accessibility, and 5mC data for a systems-level view.

At CD BioSciences, our 5hmC analysis service is designed to support your research in neurobiology, cancer epigenetics, developmental biology, environmental epigenetics, aging, and disease mechanisms. In addition to 5hmC, we also offer comprehensive analysis of other DNA modifications, including 5mC, 5fC, 5caC, 6mA, 4mC, and oxidative damage markers such as 8-oxoG. Our flexible and customizable workflows allow you to tailor the analysis to your specific research focus. If you are interested in our services, please feel free to contact us for more details and quotation information of related services.

Reference
1. Ecsedi S, Rodríguez-Aguilera J R, Hernandez-Vargas H. 5-Hydroxymethylcytosine (5hmC), or how to identify your favorite cell[J]. Epigenomes, 2018, 2(1): 3.