Reduced Representation Bisulfite Sequencing (RRBS) offers a cost-effective, focused solution to perform genome-scale DNA methylation analysis at the single nucleotide level in any vertebrate species. The fundamental idea of RRBS is to get a “reduced representation” of the genome. By cutting the genome using the restriction MspI enzyme (CCGG target sites) followed by size selection, the DNA sample is enriched with biologically relevant CpG-rich regions (including promoters and CpG islands) in which DNA methylation marks are typically found.
RRBS workflow allow you to:
- Perform cost-effective and reliable genome-scale DNA methylation analysis
- Detect- up to 4 million CpGs (with coverage > 10) in human samples
- Identify methylation patterns in CpG rich regions including promoters and CpG islands
- Benefit from increased accuracy with the use of Unique Molecular Identifiers (UMIs)
- Get better insight of your epigenetics research
Diagenode's Premium RRBS kit V2 has been further optimized to generate RRBS libraries from the lowest DNA amounts (down to 25 ng) and secure high-quality NGS data for DNA methylation analysis. Combined with unique molecular identifiers (UMI), this complete solution ensures an improved quantification accuracy during the DNA methyaltion analysis.
Premium RRBS kit V2 includes all reagents necessary for the library preparation. Specific adapters were designed and validated to fit the Premium technology and are available separately:
For RRBS UDI-UMI library construction:
How Do UMIs Work?
UMIs incorporate a unique barcode onto each molecule within a given RRBS library. By incorporating individual barcodes on each original DNA fragment, variant alleles present in the original DNA sample (true variants) can be distinguished from errors introduced during library preparation or sequencing.
Figure 1. Premium RRBS V2 construct. Integrated UMIs enable removal of PCR duplicates during the data analysis.
Accurate Coverage of CpGs
Table 1. Examples of Premium RRBS V2 sequencing data. UMI data processing enables accurate estimation of CpG counts.