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RRBS Service (Reduced Representation Bisulfite Sequencing)

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  •  Description
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Catalog Number
Format
G02020000

RRBS Service

Reduced representation bisulfite sequencing (RRBS) enables genome-scale DNA methylation analysis at the single nucleotide level in any vertebrate species. The assay benefits from the practical advantages of bisulfite sequencing while avoiding the cost of whole genome sequencing. By cutting the genome using the restriction MspI enzyme (CCGG target sites) followed by size selection, DNA is enriched to represent biologically relevant target CpG-rich regions including promoters and CpG islands. Our RRBS service makes this technology widely available and provides high coverage (up to 5 million CpGs detected in human samples).

Genome-scale DNA methylation analysis

  • Robust and cost-effective solution with reliable results
  • High coverage - up to 5 million CpGs detected in human samples
  • Single nucleotide resolution
  • Detection of methylation patterns in CpG rich regions including promoters and CpG islands
  • Suitable for epigenetic biomarker discovery

Complete end-to-end service

  • From wet-lab to bioinformatics for only 550 per sample!
  • Leveraging our Premium RRBS kit widely adopted
  • As little as 100 ng for any vertebrate species
  • Differentially methylated site analysis using standard bioinformatics tools

Suitable for any vertebrate species

Species No. of uniquely aligned reads No. of CpGs detected No. of CpGs detected with coverage >10x Average coverage (×)
Human 24,146,680 5,423,632 2,931,645 16
Mouse 25,696,428 2,473,756 1,891,964 35
Rat 23,938,442 2,391,172 1,856,947 35
Examples of RRBS data generated with our RRBS service team on Illumina's NovaSeq platform using samples from different species

See our other DNA Methylation Profiling Services

TESTIMONIAL

We are very happy with the services provided by Diagenode. In our project, Reduced Representation Bisulfite Sequencing (RRBS) was used and it worked well for both high-quality DNA and DNA extracted from formalin-fixed, paraffin-embedded (FFPE) material. Bioinformatic analyses were comprehensive and high-quality results were obtained. Diagenode’s services can be recommended.

Satu Mäki-Nevala, PhD, University of Helsinki
  • Services Workflow

    RRBS Service includes:

    QC of the genomic DNA
    • Measurement of DNA concentration 
    • Assessment of DNA quality
    Preparation of RRBS libraries
    • MspI digestion
    • Library preparation (ends preparation, adaptor ligation)
    • Size selection
    • Sample pooling
    • Bisulfite conversion
    • Library amplification and clean-up
    • QC of the RRBS library pool (DNA concentration, analysis of the pool profile)
    Deep sequencing
    • Samples are sequenced on an Illumina platform, paired-end reads, read length 50 bp (PE50)
    • 40 million raw reads (on average) per sample when pooling 10 samples/lane
    • 3-4 million CpGs (on average) for human samples
    • 10-15x CpG coverage (on average) for human samples
  • Bioinformatics Analysis

    Analysis

    Features

    Standard
    • FASTQ raw data
    • FASTQC quality control insights
    • Alignment of bisulfite sequencing data against reference genome
    • Methylation calling and extraction
    • Summary statistics
    Differential methylation analysis
    • Methylation level analysis
    • Differentially Methylated CpGs (DMCs) analysis
    • Differentially Methylated Regions (DMRs) analysis
    • Annotation of DMCs and DMRs for genomic regions (exons, introns, …)
    • Clustering analysis

    Gene ontology terms analysis

    • Enrichment analysis on gene associated with DMCs and DMRs
    • Get functional insights

    Pathway analysis

    • Identification of biological pathways in which genes associated with DMCs and DMRs may be over-represented (or under-represented)
    • Get mechanistic insights
  •  Testimonials

    Our laboratory has used RRBS sevice of Diagenode on murine and human samples. The service was impeccable in each phase, from the sample preparation to bionformatic analysis because it was always customer-oriented. I highly recommend my colleagues to use the RRBS service from Diagenode.

    Prof. Lucia Altucci, MD, PhD, Seconda Università degli Studi di Napoli, Dipartimento di Biochimica, Biofisica e Patologia generale .

    The new Diagenode Premium RRBS Kit makes it easy to use RRBS cost-effectively and with high throughput, using early sample pooling and multiplex sequencing. Most importantly, the method provides an improved coverage of up to 4 million CpGs for the human genome. We successfully used this protocol on more than 1,000 samples comprising of six different species, various cancers, FFPE and lowinput samples.

    Paul Datlinger and Christoph Bock, CeMM Research Center for Molecular Medicine of the Austrian Academy of Sciences, Vienna, Austria

    We have had an excellent experience with the RRBS service provided by Diagenode. Our project was based on DNA extracted from human fresh and paraffin-embedded skeletal muscle, and the RRBS and Bioinformatic results were reliable and consistent with what we expécted. We highly recommend their service

    Mario Roque - Instituto de Histología y Embriología de Mendoza (IHEM), Mendoza, Argentina
  •  Documents
    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. ...
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    The Diagenode Epigenetics custom service POSTER
    Complete workflows for genome-scale DNA methylation and histone marks analysis Epigenetics is cr...
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    Epigenomics Profiling Services FLYER
    Chromatin analysis DNA methylation services RNA-seq analysis
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  •  Publications

    How to properly cite this product in your work

    Diagenode strongly recommends using this: RRBS Service (Reduced Representation Bisulfite Sequencing) (Diagenode Cat# G02020000). Click here to copy to clipboard.

    Using our products in your publication? Let us know!

    Genome-Wide Epigenomic Analyses in Patients With Nociceptive and Neuropathic Chronic Pain Subtypes Reveals Alterations in Methylation of Genes Involved in the Neuro-Musculoskeletal System
    Stenz et al
    Nociceptive pain involves the activation of nociceptors without damage to the nervous system, whereas neuropathic pain is related to an alteration in the central or peripheral nervous system. Chronic pain itself and the transition from acute to chronic pain may be epigenetically controlled. In this cross-sectional s...

    Genome-wide epigenomic analyses in patients with nociceptive and neuropathic chronic pain subtypes reveals alterations in methylation of genes involved in the neuro-musculoskeletal system
    Ludwig Stenz, Joane LeCarré, François Luthi, Philippe Vuistiner, Cyrille Burrus, Ariane Paoloni-Giacobino, Bertrand Léger
    Nociceptive pain involves the activation of nociceptors without damage to the nervous system, whereas neuropathic pain is related to an alteration in the central or peripheral nervous system. Chronic pain itself and the transition from acute to chronic pain may be epigenetically controlled. In this cross-sectional s...

    Environmental enrichment preserves a young DNA methylation landscape in the aged mouse hippocampus
    Sara Zocher, Rupert W. Overall, Mathias Lesche, Andreas Dahl & Gerd Kempermann
    The decline of brain function during aging is associated with epigenetic changes, including DNA methylation. Lifestyle interventions can improve brain function during aging, but their influence on age-related epigenetic changes is unknown. Using genome-wide DNA methylation sequencing, we here show that experiencing ...

    Integrative Analysis of Glucometabolic Traits, Adipose Tissue DNA Methylation and Gene Expression Identifies Epigenetic Regulatory Mechanisms of Insulin Resistance and Obesity in African Americans
    Neeraj K. Sharma, Mary E. Comeau, Dennis Montoya, Matteo Pellegrini, Timothy D. Howard, Carl D. Langefeld, Swapan K. Das
    Decline in insulin sensitivity due to dysfunction of adipose tissue (AT) is one of the earliest pathogenic events in Type 2 Diabetes. We hypothesize that differential DNA methylation (DNAm) controls insulin sensitivity and obesity by modulating transcript expression in AT. Integrating AT DNAm profiles with transcrip...

    Chronic cannabidiol alters genome-wide DNA methylation in adult mouse hippocampus: epigenetic implications for psychiatric disease.
    Wanner NM, Colwell M, Drown C, Faulk C
    Cannabidiol (CBD) is the primary non-psychoactive compound found in cannabis (Cannabis sativa) and an increasingly popular dietary supplement as a result of widespread availability of CBD-containing products. CBD is FDA-approved for the treatment of epilepsy and exhibits anxiolytic, antipsychotic, prosocial, and oth...

    LY75 Ablation Mediates Mesenchymal-Epithelial Transition (MET) in Epithelial Ovarian Cancer (EOC) Cells Associated with DNA Methylation Alterations and Suppression of the Wnt/β-Catenin Pathway.
    Mehdi S, Bachvarova M, Scott-Boyer MP, Droit A, Bachvarov D
    Growing evidence demonstrates that epithelial-mesenchymal transition (EMT) plays an important role in epithelial ovarian cancer (EOC) progression and spreading; however, its molecular mechanisms remain poorly defined. We have previously shown that the antigen receptor LY75 can modulate EOC cell phenotype and metasta...

    Obesogen effect of bisphenol S alters mRNA expression and DNA methylation profiling in male mouse liver
    Brulport Axelle, Vaiman Daniel, Chagnon Marie-Christine, Le Corre Ludovic
    Environmental pollution is increasingly considered an important factor involved in the obesity incidence. Endocrine disruptors (EDs) are important actors in the concept of DOHaD (Developmental Origins of Health and Disease), where epigenetic mechanisms play crucial roles. Bisphenol A (BPA), a monomer used in the man...

    Demethylation of ITGAV accelerates osteogenic differentiation in a blast-induced heterotopic ossification in vitro cell culture model.
    Logan NJ, Camman M, Williams G, Higgins CA
    Trauma-induced heterotopic ossification is an intriguing phenomenon involving the inappropriate ossification of soft tissues within the body such as the muscle and ligaments. This inappropriate formation of bone is highly prevalent in those affected by blast injuries. Here, we developed a simplified cell culture mod...

    Paternal sepsis induces alterations of the sperm methylome and dampens offspring immune responses—an animal study
    Katharina Bomans, Judith Schenz, Sandra Tamulyte, Dominik Schaack, Markus Alexander Weigand and Florian Uhle
    Abstract Background Sepsis represents the utmost severe consequence of infection, involving a dysregulated and self-damaging immune response of the host. While different environmental exposures like chronic stress or malnutrition have been well described to reprogram the germline and subsequently offspring a...

    DNMT3B overexpression contributes to aberrant DNA methylation and MYC-driven tumor maintenance in T-ALL and Burkitt’s lymphoma
    Poole et al.
    Aberrant DNA methylation is a hallmark of cancer. However, our understanding of how tumor cell-specific DNA methylation patterns are established and maintained is limited. Here, we report that in T-cell acute lymphoblastic leukemia (T-ALL) and Burkitt’s lymphoma the MYC oncogene causes overexpression of DNA me...

    Diagenode® Premium RRBS technology: cost-effective DNA methylation mapping with superior coverage
    Anne-Clémence Veillard, Paul Datlinger, Miklos Laczik, Sharon Squazzo & Christoph Bock
    Reduced representation bisulfite sequencing (RRBS) enables genome-scale DNA methylation analysis in any vertebrate species. The assay benefits from the practical advantages of bisulfite sequencing while avoiding the cost of whole-genome sequencing. The Diagenode Premium RRBS kit makes this technology widely availabl...

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Events

  • DNA methylation workshop: Become an expert at RRBS
    Diagenode
    Oct 1-Oct 31, 2021
  • Dutch Chromatin Meeting
    Radboud UMC, Geert Grooteplein 15, 6525 EZ Nijmegen, The Netherlands, Experience center, route 292 to 307
    Nov 12, 2021
 See all events

News

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