A single-cell resolution DNA methylation analysis technique is required when conducting studies on the DNA methylation analysis of trace or heterogeneous cells such as embryonic stem cells and tumor stem cells. Creative BioMart offers the single-cell whole-genome bisulfite sequencing (scWGBS) service, which is capable of performing single-cell DNA methylation analysis in a wide range of biological systems to explain intercellular epigenetic heterogeneity in complex tissue samples.

What Is Single-cell WGBS?

The single-cell whole-genome bisulfite sequencing (scWGBS) is a methylome analysis technique that offers single-base resolution and absolute quantification of 5mC genome-wide within an individual cell. Compared with the single-cell reduced representation bisulfite sequencing (scRRBS) focusing on CpG islands, scWGBS is relatively unbiased and provides cumulative coverage for more than 90% of CpG loci in the human and mouse genomes. Incorporating epigenetic information into a single cell library will transform our understanding of gene regulation and provide insights into epigenetic heterogeneity for a wide range of applications, such as epigenomic analysis of heterogeneous organs and epigenetic drug resistance research.

In the commonly utilized protocol of whole-genome bisulfite sequencing (WGBS), sequencing adapters are ligated to the fragmented DNA prior to bisulfite conversion, which results in the loss of information due to the degradation of the DNA treated by bisulfite. To minimize DNA loss in individual cells, Creative BioMart adopts the strategy of post-bisulfite construction, where bisulfite conversion takes place prior to addition of sequencing adapters. Additionally, Creative BioMart can sort the cells used for DNA methylation analysis by fluorescence-activated cell sorter (FACS) according to customers’ needs.

Fig 1. The schematic diagram of scWGBS

Features of Single-cell WGBS

1. Cumulative coverage for more than 90% of CpG loci across the entire genome
2. Accurately measure 5mC across genomes of single cells
3. Flexible and enables DNA methylation mapping both in very small cell populations and single cells

Our Advantages

1. Skillful scientific team: we will offer you with our superior technical support for your research, which will be motivated by a wealth of scientific publications as well as our in-house experience.
2. Unsurpassed data quality: we guarantee that more than 80% of bases with a ≥Q30 quality score, exceeding Illumina’s official guarantee of ≥ 75%.
3. Comprehensive data analysis: we use widely-accepted mainstream software such as Bismark and a mature in-house pipeline for mapping, DMR analysis, functional analysis, and data visualization, providing you with both standard data analyses and customized bioinformatics analyses.

Workflow of Single-cell WGBS at Creative BioMart
Creative BioMart has established a fast and cost-effective scWGBS workflow to handle large numbers of samples. Our one-stop service includes single cell isolation, bisulfite conversion, library construction, and next-generation sequencing. We also provide data analysis service including alignment and methylation status calling at both single CpG resolution and regional resolution for scWGBS.

Fig 2. Workflow of scWGBS at Creative BioMart

Please feel free to contact us with your projects associated with DNA methylation analysis of trace or heterogeneous cells. Creative BioMart is here to offer you professional and thoughtful service!

References
1. Smallwood S A.; et al. Single-Cell Genome-Wide Bisulfite Sequencing for Assessing Epigenetic Heterogeneity. Nature Methods. 2014, 11(8): 817.
2. Farlik M.; et al. Single-Cell DNA Methylome Sequencing and Bioinformatic Inference of Epigenomic Cell-State Dynamics. Cell Reports. 2015, 10(8): 1386-1397.