Compared to whole genome bisulfite sequencing (WGBS), reduced representation bisulfite sequencing (RRBS) provides an alternative way to generate and profile comprehensive methylomes at a reduced cost. RRBS combines restriction enzymes and bisulfite sequencing to enrich CpG rich genomic regions for methylation state detection. The methylation insensitive restriction enzyme MspI is commonly used in RRBS to cut genomic DNA at CCGG sites. After end-repair, A-tailing, adapter ligation of the fragmented genomic DNA, the library is then size-selected for fragments with length between 40 and 220bp. This will reduce the amount of base pairs to be sequenced down to 1~2.5% of the genome but still keep vast majority of promoter sequences and CpG islands (CGIs). Then these CpG content enriched fragments are bisulfite converted and sequenced same as WGBS.
Technique Highlights
－High Resolution Methylation state detection at single-nucleotide resolution
－Cost Effective Sequencing only GpC-enriched genomic regions dramatically decreases the required volume of sequencing data as well as the cost
－Informative representation of genome-wide methylome over 85% of CpG islands and 60% of promoters are captured and analyzed in RRBS
－Allow methylation detection in repeated sequences repeated sequences often contain methylated cytosines and changes of methylation patterns in these repeated sequences are often associated with cancer pathogenesis. RRBS provides an effective mean for methylation detection in these challenging regions which are not accessible by current sequencing technologies.
Sample Requirement
Sample Type: Genomic DNA
Amount: DNA >= 5 ug; concentration >= 100 ng/ul
Purity: OD260/280 = 1.7~2.0
Sample without severe degradation
Data generation criteria
Sequencing is conducted by strictly following the manufacturer’s SOP of cBot and HiSeq. Sequencing mode 2 X 125bp, index 8bp.
Service Workflow
1. Customized experimental design
Deliverables
