InquiryDuring normal cell development, DNA methylation serves as an important "epigenetic marker" or a chemical or protein tag on the DNA sequence. Without altering the existing DNA sequence, this tag keeps track of cellular changes and participates in the biological regulation of processes that turn on or off, controlling the cellular life cycle.
In fact, the sequences of genomic DNA of different cell types of the same organism are almost identical. DNA molecules of the same sequence may contain methylation modifications, and these various modifications are the foundation of different cell types; meanwhile, cells of the same type may also preserve specific gene expression characteristics. As a result, these different modifications and modification combinations allow the formation of heterogeneity between cells of the same type. Therefore, genome methylation modification specificity at the single-cell level is an important research application for epigenetics.
Single-cell whole genome bisulfite sequencing (scWGBS) is a technique to study whole genome methylation using the heavy sulfite treatment at the single-cell level combined with the high-throughput Illumina sequencing platform. This approach precisely resolves the methylation status of each cytosine on the single-cell level and preserves significant implications for uncovering epigenetic information in highly heterogeneous cells. Nevertheless, this technique offers the possibility to reveal the developmental mechanisms of embryos, cancer, and infertility research treatments.
Workflow of scWGBS Service
Technical Advantages
Cells were first treated with BS, followed by adapter amplification to minimize the loss of DNA methylation modifications in single cells.
The addition of adapters during random primer amplification can reduce operation steps and prevent DNA loss.
Bioinformatics Analytical Contents
| Statistical data output and sequencing quality |
| Compare reads in Clean Data of the reference genome and count alignment rates, duplication rates, insert size distributions, and average sequencing depths |
| Count C site methylations of the whole genome, BS conversion rates, and in-depth coverage of the C sites |
| C site coverage of CGI and promoter regions |
| Distribution of methylation levels |
| The number of methylated/unmethylated sites |
| Methylation status of CGI |
| Sample clustering |
| Analysis of homogeneity/heterogeneity |
| Calculation and annotation of DMR area between groups |
| Methylated/unmethylated regions |
Sample Requirements
Samples can be submitted as single cells (nuclei) or frozen cells in cell suspensions
No upper limit on the number of fresh cells; cell suspensions < 4 μL
Sample requirement: > 3 single-cell replicates are recommended
Species: human, mouse or other mammalian (high-quality reference genome required)
Please for more sample requirement details.
Applications
Study of genetic recombination patterns for germ cells
Embryo development studies
Cell heterogeneity studies
Cancer research
FAQ
How to choose between single-cell targeted region methylation and single-cell whole genome methylation?
Single-cell targeted region methylation (ScRRBS) is a co-enzymatic method that enriches the CpG island regions, mainly detecting the methylation status of the promoter regions of genes. Meanwhile, single-cell whole genome methylation (ScWGBS) sequences obtain in-depth genome-wide methylation status information.
