Reduced Representation Bisulfite Sequencing (RRBS)

Input Requirement(s)
250ng of genomic DNA at a concentration of 10ng/ul

Batch Size: 24 samples or 84 samples

Bisulfite sequencing is a powerful approach to mapping of methylated cytosine residues at single base resolution. We offer a targeted or ‘Reduced Representation Bisulfite Sequencing’ (RRBS) method which enables a lower cost approach than whole genome bisulfite sequencing, while providing enriched coverage of CpGs. Our RRBS method (Meissner et al., 2005; Gu et al., 2011) utilizes a methylation-insensitive restriction enzyme (MspI) to generate short DNA fragments with CpG dinucleotides at their ends, which are then subjected to bisulfite conversion, PCR amplification and Illumina library construction. Selectively targeting CG-rich regions of the genome in this manner allows for a significant reduction in the amount of sequencing required while still capturing the majority of promoters and other genomic regions of interest. This approach is highly sensitive, providing quantitative DNA methylation readout. With RRBS, approximately 10% of all CpG dinucleotides in the human genome can be profiled. This method can also be applied to murine and other suitable genomes.