InquiryAs a cost-effective methylation study method, Single-cel Reduced Representation Bisulfite Sequencing (ScRRBS) has broad application prospects in basic research and large-scale clinical sample research.
Epigenetics refers to changes in gene expression levels based on non-genetic sequence changes, such as DNA methylation and chromatin conformational changes. Epigenetics is the study of reversible, heritable changes in gene function in the absence of alterations in the DNA sequence of the cell nucleus. Epigenetic modifications, which regulate gene activity without altering DNA sequences, have profound implications for human development, human disease, and environmental impacts. DNA methylation is one of the most common epigenetic modifications that is widely involved in cellular control of gene expression and plays a critical role in cell growth, cell differentiation, cell proliferation and disease states. DNA methylation is an important epigenetic marker, closely related to gene expression and phenotypic traits, and has been a hot spot for epigenetic research. Detection of epigenetic status by single cell analysis is a major trend in single cell research.
Reduced Representation Bisulfite Sequencing (RRBS) is an accurate, efficient, and cost-effective method for DNA methylation studies, which enriches promoter and CpG island regions within the whole genome by specific enzymatic cleavage and Bisulfite sequencing to high-resolution detect DNA methylation. This method can obtain CpG site information of some important regions, including more than 70% of promoter regions and more than 80% of methylation islands.
Our Single-cell Methylation Detection Solution
CD Genomics has established a series of micro- and single-cell methylation detection methods, which can provide personalized detection solutions for different project requirements, and carry out methylation detection of whole genome, and target genes. We also offer comprehensive single-cell epigenetics analysis services, such as Single-cell Whole Genome Bisulfite Sequencing and Single-cell ATAC Sequencing
Single-cell RRBS Workflow
Genome Research 2013; 23(12)
Sample Requirements
1. Isolation of single cells from tissue requires fresh tissue without freezing, fixation, or other processing.
2. Ensure cell viability and morphological integrity.
3. Dissociated single cells were placed in lysis buffer using 200 µl PCR tubes. Each tube of lysate is 5µl, and the volume of buffer should not exceed 1µl.
4. Number of cells: One or more cells are placed in each tube, and 3-5 repetitions are recommended for each cell.
5. Single cells were placed in lysis buffer and stored at -80°C and shipped on dry ice.
6. Keep the low temperature environment as much as possible during the operation.
7. The lysate added with single cells should be stored at -80°C as soon as possible. It is best not to exceed 1 hour. The shorter the intermediate time, the better.
8. Once the sample has been frozen at -80°C or on dry ice, it cannot be thawed repeatedly.
9. Species: human, mouse or other mammalian (high-quality reference genome required). Please for more sample requirement details.
FAQ
How to choose between single cell restricted region methylation and single cell whole genome methylation?
Single-cell Reduced Representation Bisulfite Sequencing (ScRRBS), which is a co-enzymatic method that enriches more of the CpG island region, is primarily used to detect the methylation status of the promoter region of the gene.
Single-cell whole genome bisulfite sequencing (ScWGBS)is to lyse cells, digest the proteins that encapsulate DNA, treat the DNA with sulfite, followed by several rounds of linear amplification with random primers so that the region obtained is random on the genome, and then performing exponential amplification so that the whole genome can be obtained if the sequencing is deep enough.
What are the applications of Single cell RRBS?
It mainly detects the methylation status of the promoter region of the gene.
