Introduction
Whole genome hydroxymethylated DNA co immunoprecipitation sequencing (hydroxymethylated DNA)
Immunoprediction sequencing (hmedip SEQ) uses 5 '- hydroxymethylcytosine specific antibody to enrich hydroxymethylated DNA fragments, and then combines with high-throughput sequencing technology to quickly and efficiently search for hydroxymethylated regions at the whole genome level with a small amount of data. It can be widely used in the study of the relationship between hydroxymethylation and disease and the study of hydroxymethylation in the process of embryonic development
Technical advantages
1.Applywide range
Suitable for people and most animals and plants
2. genome wide coverage
Maximize access to complete genome-wide methylation information and accurately map methylation
3. single base resolution
It can accurately analyze the methylation status of each C base
Experimental strategy
Information analysis
Technical parameter
| Sample requirements | Project cycle |
| Total amount: different samples have different requirements, human samples ≥ 2 μ g; Purity: od260/280 = 1.8~2.0; Integrity: there is no obvious degradation of DNA, and gel electrophoresis detection gel map shall be provided. Recommended data volume: 4G clean data | The operation cycle of the standard process below 15 samples is about 30 working days. When the number of samples is large, the project cycle needs to be determined with the technical support personnel. |
Case analysis
Case: hydroxymethylation of mouse embryonic stem cells
Wu H, et al. Genome-wide analysis of 5-hydroxymethylcytosine distribution reveals its dual function in transcriptional regulation in mouse embryonic stem cells. Genes Dev. 2011 Apr 1;25(7):679-84.
Background：
Studies have confirmed that tet family proteins can catalyze the conversion of 5mC to 5hmc. 5mC has been widely studied, but little is known about the distribution and function of 5hmc.
Methods：
Hmedip SEQ was used to study the wild-type and Tet1 depleted 5hmc of mouse embryonic stem cells.
Conclusion:
5hmc is located in the gene bodies region of genes with high transcriptional activity and the promoter region of developmental regulators repressed by Polycomb; 5hmc may play dual functions of activation and repression in the regulation of gene expression.
Genome wide distribution of 5hmc
