Premium RRBS kit V2
RRBS for low DNA amounts and accurate analysis

Catalog Number
24 rxns

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. 

With Diagenode’s Premium RRBS Kit V2 perform cost-effective and reliable genome-scale DNA methylation analysis. Detect around 4 million CpGs (with coverage > 10) in human samples and identify methylation patterns in CpG rich regions including promoters and CpG islands

Secure high-quality NGS data for DNA methylation analysis at single base resolution and enjoy our specific and upgraded features:

  • Work with the lowest DNA amounts from 25 -100ng gDNA
  • Process up to 96 sample in parallel while saving handling-time and cost per sample with early sample pooling strategy
  • Utilize our Software for Intelligent Pooling (SIP) for better pooling strategies
  • Get optimal results from your sequencer with Unique Dual Indexing (UDI)
  • Identify and remove PCR duplicates from your data with Unique Molecular Identifiers (UMIs)

The Premium RRBS kit V2 includes all reagents necessary for the library preparation. Specific adapters were designed and validated to fit the technology and are available separately as described below.

For RRBS V2 with UDI and 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.

  • Validation

    Sequencing quality



    Figure 2. Excellent sequencing quality. RRBS libraries were prepared from different starting amounts of human gDNA using Diagenode’s Premium RRBS V2 kit and sequenced in paired-end 50 bp on Illumina NovaSeq instrument generating 30-40 million read pairs per sample. Sequencing statistics reveal that all samples performed well with mean Phred scores above 30 along the entire reads 1 (A) and 2 (B) (data shown for 50 ng gDNA input after trimming).

    Accurate Coverage of CpGs

    Table 1. Examples of Premium RRBS V2 sequencing data. UMI data processing enables accurate estimation of CpG counts.

    Focus on CpG-rich regions

    Figure 3. Coverage of CpGs and genomic regions by Premium RRBS V2Diagenode’s Premium RRBS V2 allows a wide interrogation of CpGs (with a sequencing depth >10) of the human genome with a focus on CpG rich regions, especially CpG islands (data shown for 50 ng gDNA input).

  • Nature Method & Videos


    Premium RRBS technology: cost-effective DNA methylation mapping with superior coverage

    Nature Methods 13 (2016)

    Published online


  • Software for Intelligent Pooling

    Diagenode's new online intelligent pooling aid (RRBS SIP) provides the optimal pool design for RRBS to meet your specific sample and analysis needs:

    • Time-saving:  avoid complex caculations
    • Highest pooling efficiency based on qPCR quantification - elevate your pooling effectiveness
    • Powerful: incorporates advanced aspects such as number of samples per pool required and the separation between projects
    • Accurate: identifies outliers

    Get access to Diagenode's RRBS SIP - coming soon.

    Get your RRBS template file - Here

  •  Documents
    Template RRBS Pooling file EXCEL FILE
    Template CSV file for RRBS pooling tool
    Premium RRBS spike-in controls manual MANUAL
    This document and files are to process the sequencing data from the spike-in controls included in...
    Sequence of the methylated spike-in control in FASTA format
    This document and files are to process the sequencing data from the spike-in controls included in...
    Sequence of the unmethylated spike-in control in FASTA format
    This document and files are to process the sequencing data from the spike-in controls included in...
    Positions of the unmethylated cytosines in the sequence of the methylated control in BED format
    This document and files are to process the sequencing data from the spike-in controls included in...
    DNA Methylation Solutions BROCHURE
    Complete solution for DNA methylation studies! Whether you are experienced or new to th...
    Diagenode Premium RRBS technology: Cost-effective DNA methylation mapping with superior CpG resolution and coverage APPLICATION NOTE
    DNA methylation is an important epigenetic mark with broad relevance in development and disease. ...
    Premium RRBS kit v2 manual MANUAL
    Manual Premium RRBS kit V2
  •  Publications

    How to properly cite this product in your work

    Diagenode strongly recommends using this: Premium RRBS kit V2
    RRBS for low DNA amounts and accurate analysis (Diagenode Cat# C02030036)
    . Click here to copy to clipboard.

    Using our products in your publication? Let us know!

    Multi-omics phenotyping of the gut-liver axis reveals metabolicperturbations from a low-dose pesticide mixture in rats.
    Mesnage, Robin et al.
    Health effects of pesticides are not always accurately detected using the current battery of regulatory toxicity tests. We compared standard histopathology and serum biochemistry measures and multi-omics analyses in a subchronic toxicity test of a mixture of six pesticides frequently detected in foodstuffs (azoxystr...

    The insecticide permethrin induces transgenerational behavioral changeslinked to transcriptomic and epigenetic alterations in zebrafish (Daniorerio).
    Blanc, Mélanie et al.
    The pyrethroid insecticide permethrin is widely used for agricultural and domestic purposes. Previous data indicated that it acts as a developmental neurotoxicant and can induce transgenerational effects in non-target organisms. However, associated underlying mechanisms remain unclear. The aim of this study was to i...

    Hepatic transcriptome and DNA methylation patterns following perinataland chronic BPS exposure in male mice.
    Brulport A. et al.
    BACKGROUND: Bisphenol S (BPS) is a common bisphenol A (BPA) substitute, since BPA is virtually banned worldwide. However, BPS and BPA have both endocrine disrupting properties. Their effects appear mostly in adulthood following perinatal exposures. The objective of the present study was to investigate the impact of ...

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