RRBS (Reduced Representation Bisulfite Sequencing)
single base pair resolution DNA methylation analysis
Reduced Representation Bisulfite Sequencing (RRBS) provides DNA methylation data at single base pair resolution on up to 5 million CpGs. More cost effective than Whole Genome Bisulfite Sequencing and with greater coverage and less bias than bead array-based DNA methylation platforms, RRBS is an ideal platform to profile DNA methylation, with a focus on functional loci, genome-wide.
- DNA methylation data on up to 5 million CpGs per sample.
- Biologically relevant target regions including promoters and CpG islands.
- Single-end sequencing depth of >30,000,000 reads.
- Low input DNA requirement of 100 ng.
- Reduced library bias through from removal of PCR duplicates by random bar code incorporation.
- Raw data (FASTQ) output files
- Aligned data (BAM) files
- Methylation table includes percent methylation value for each CpG
- Differential methylation analysis
- Figures & graphs
– Includes coverage depth analysis, correlation plots, and pie charts
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RRBS Background & Data
RRBS is a method designed to isolate DNA containing CpGs by using DNA restriction endonuclease digestion and amplification of fragments smaller than 300 bp. In Brief, the methylation insensitive endonucleases, Msp1 and Taq1, are used to cut C^CGG and T^CGG respectively, thus generating DNA fragments with CpG at the end. Since CpG islands and promoters have a high density of CpGs they are cut at a high frequency and the fragments derived from these regions tend to be short. This subset of short fragments is preferentially amplified when the library is generated through PCR and additional selection occurs during sequencing cluster generation. The final fraction of DNA that is sequenced represents a small fraction of the total genome and is enriched in CpGs. These fragments are mapped to the genome to reveal the methylation status of millions of individual CpGs.