DNA hydroxymethylation is an important epigenetic modification that regulates gene expression and plays an important role in neural differentiation and occurrence and development of cancer. To understand the role of 5hmC, we must understand the distribution of 5hmC in the genome. However, conventional bisulfite-based methods cannot distinguish between 5hmC and 5mC. At Creative BioMart, we combine the 5hmC monoclonal antibody capture approach with high-throughput sequencing and bioinformatics analysis to obtain a genome-wide hydroxymethylation profile, thus helping customers to analyze the molecular mechanism of embryonic development, neural cell differentiation and cancer development from a new perspective.

What Is hMeDIP-Seq?

Consistent with the principle of genome-wide methylated DNA immunoprecipitation sequencing (MeDIP-Seq), hydroxymethylation DNA immunoprecipitation sequencing (hMeDIP-Seq) separates hydroxymethylated DNA from genomic DNA through immunoprecipitation. Antibodies against 5hmC are incubated with the fragmented genomic DNA and precipitated, followed by DNA purification and library preparation. Deep sequencing provides greater genomic coverage, representing the majority of the immunoprecipitated hydroxymethylated DNA. The hMeDIP-Seq platform efficiently and accurately detects 5-hydroxymethylcytosine (5hmC) in the entire genome at multi-base resolution (50-100 bp) without cross-reaction with methylated cytosine or unmethylated cytosine. 5hmC antibodies of high affinity are utilized to immunoprecipiate and enrich DNA fragments with the highest accuracy.

Figure 1. The schematic diagram of hMeDIP

Features of hMeDIP-Seq

1. Affinity-based capture approach, the high sensitivity and specificity of antibodies are utilized
2. Cover 5hmC in dense and low-density repeating areas
3. Specific to 5hmC depending on the antibody specificity
4. Cost-effective technique

Our Advantages

1. Tolerance of multiple sample types: we currently accept a wide range of biological and clinical samples including cells and tissues.
2. Flexible and one-stop service: customers only need to provide samples, Creative BioMart complete the whole service process from DNA sample preparation, hMeDIP enrichment, library preparation, on-machine sequencing to data analysis.
3. Strict quality control: Creative BioMart adds quality control (QC) at every critical step in hMeDIP-Seq. These QC data can assess the quality of each step. If the standard is not reached, we will repeat the experimental steps or optimize the experimental system, so that each sample can smoothly enter the next experimental stage.
4. Professional bioinformatics analysis: our professional bioinformatics analysis tools, such as MACS and diffReps, can help you accomplish hydroxymethylation peaks identification, differential hydroxymethylation gene recognition, cluster analysis / GO analysis / Pathway analysis of differential hydroxymethylation genes. The visualized map provided by us enables you to visually understand the hydroxymethylation of specific regions or genes, as well as the differential hydroxymethylation status between samples.

Workflow of hMeDIP-Seq at Creative BioMart

Creative BioMart provides you with hMeDIP-Seq service, you only need to provide well-preserved tissue or cell samples, our technical staff can complete all the experimental operations, including DNA sample preparation (sonication, ligation of sequencing adapters and denaturation), hMeDIP enrichment, input DNA library construction, high-throughput sequencing, as well as data analysis.

Figure 2. Workflow of hMeDIP-Seq at Creative BioMart

Creative BioMart’s breadth of experience gained from being in the industry for more than 10 years makes us an expert partner for your epigenetics projects. contact us and start your project.

Reference
1. Lian.; et al. Loss of 5-hydroxymethylcytosine is an epigenetic hallmark of melanoma. Cell. 2012, 150(6): 1135-1146.