Diagenode

Premium RRBS kit (24 rxns) - replaced by Premium RRBS kit v2 C02030036

Catalog Number
Format
Price
C02030032
24 rxns
Other format
DISCONTINUED

The kit Premium RRBS (24 rxns) has been replaced by an upgraded version: Premium RRBS v2 (24 rxns). Please note, that the adapters are not included in the kit and must be purchased separately. For more information please check out: Premium RRBS v2 (24 rxns). 
For the information about the format 96 rxns - check out Premium RRBS (96 rxns)

  • Characteristics
    • Superior coverage – 4 million CpGs
    • High throughput - 96 samples processed in one experiment
    • Cost-efficient – Multiplex up to 6 samples/sequencing lane
    • Validated with FFPE, cancer, and low-input samples
    • High efficiency and minimal bias - Low DNA degradation and reduced amplification
    • Complete kit – Bisulfite conversion reagents, MspI enzyme, library preparation reagents including barcodes, and spike-in controls
    • Already tested on various species – human, mouse, rat, pig, cow, dog, zebrafish, Daphnia and more cichlid fish (Astatotilapia calliptera), mossy frog, yellow-bellied slider, dice snake, zebra finch, humboldt penguin, leaf bird, buzzard, vulturine guinea fowe, parma kangaroo, cheetah, mouflon

     

    Chr Shearing


    Superior coverage

    Comparison of CpG coverage between competing technologies.

    They love it!
    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

     

    dna methylation

    Accurate determination of DNA methylation level

    Excellent results were obtained using Diagenode’s Premium RRBS kit: almost 90% alignment rate, 4.1 million CpGs covered and bisulfite conversion rates around 99.5% for all samples. DNA methylation percentages in the region of IGF2 obtained with the Diagenode’s Premium RRBS Kit. Two human cell lines were analyzed: Gm12878 and MCF7. The MCF7 cell line was studied in duplicates. Each peak represents the DNA methylation percentage at one CpG. The methylated CpGs are shown in red and the unmethylated CpGs in grey.



    rrbs how it works

    How it works

    By cutting the genome using the restriction enzyme MspI (CCGG target sites) followed by size selection, DNA is enriched to represent CpG-rich genomic regions (including CpG islands, CpG island shores, enhancers, and other gene-regulatory elements), which are particularly relevant for epigenetic regulation. Similar to exome-sequencing for mutation discovery, the RRBS protocol enriches for some of the most interesting target regions and thereby achieves a reduction in sequencing cost of a factor of 10-20 compared to whole genome bisulfite sequencing.
     

    Premium RRBS Kit

    Illumina® RRBS protocol
    DNA input 100 ng 2-5 µg
    Multiplexing Pooling of 6 samples (one HiSeq lane) no guidelines
    Bisulfite conversion One bisulfite reaction per pool (6 samples) One bisulfite reaction per sample
    # purification steps 2 7
    Key features low input - cost-effective - optimized for high-throughput - complete kit high input - not optimized for high-throughput - reagents from different providers
  • Nature Methods

    NATURE METHODS

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

    Nature Methods 13 (2016)

    Published online

    CHECK OUT THE NATURE METHODS PAPER

  •  Testimonials

    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

    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 .

    Using the Premium RRBS kit we quickly established the RRBS library preparation in our group. Diagenode's support on wet-lab matters as well as on bioinformatics was exceptionally customer-oriented and close, accelerating the establishment of the method.

    Dr. Eduard Resch, Fraunhofer Insitute for Molecular Biology and Applied Ecology IME, Project Group Translational Medicine & Pharmacology TMP, Frankfurt am Main

    Our lab has used Diagenode’s Premium RRBS kit on rat brain samples. The protocol is understandable, logical, well-written and is easy to follow. It is a complicated, but ingenious kit. I found it fantastic that I could ask questions from the company, and their answers were really useful. We were able to construct a library, which we ran on BioAnalyzer, and the results looked very nice and ready to be sequenced. I would definitely recommend my colleagues to use the Premium RRBS kit from Diagenode.

    Borbála Vető, Institute of Enzymology, Budapest, Hungary
  •  Documents
    RRBS Pooling aid EXCEL FILE
    RRBS Pooling aid
    Download
    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. ...
    Download
    Premium RRBS kit MANUAL
    Diagenode’s Premium RRBS technology Positive and negative spike-in controls are...
    Download
    Premium RRBS spike-in controls - data processing MANUAL
    This document and files are to process the sequencing data from the spike-in controls included in...
    Download
    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...
    Download
    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...
    Download
    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...
    Download
    Premium RRBS kit V2 - Spike-in controls analysis to estimate bisulfite conversion efficiency MANUAL
    This document describes how to process demultiplexed RRBS V2 sequencing data to extract the infor...
    Download
  •  Safety sheets
    Premium Reduced Representation Bisulfite Sequencing RRBS Kit SDS US en Download
    Premium Reduced Representation Bisulfite Sequencing RRBS Kit SDS GB en Download
    Premium Reduced Representation Bisulfite Sequencing RRBS Kit SDS BE fr Download
    Premium Reduced Representation Bisulfite Sequencing RRBS Kit SDS FR fr Download
    Premium Reduced Representation Bisulfite Sequencing RRBS Kit SDS ES es Download
    Premium Reduced Representation Bisulfite Sequencing RRBS Kit SDS DE de Download
    Premium Reduced Representation Bisulfite Sequencing RRBS Kit SDS JP ja Download
    Premium Reduced Representation Bisulfite Sequencing RRBS Kit SDS BE nl Download
  •  Publications

    How to properly cite this product in your work

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    Using our products in your publication? Let us know!

    The ETS transcription factor ERF controls the exit from the naïve pluripotent state in a MAPK-dependent manner
    Maria Vega-Sendino et. al.
    The naïve epiblast transitions to a pluripotent primed state during embryo implantation. Despite the relevance of the FGF pathway during this period, little is known about the downstream effectors regulating this signaling. Here, we examined the molecular mechanisms coordinating the naïve to primed transit...

    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...

    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 ...

    Riluzole Administration to Rats with Levodopa-Induced Dyskinesia Leads to Loss of DNA Methylation in Neuronal Genes
    Luca Pagliaroli, Abel Fothi, Ester Nespoli, Istvan Liko, Borbala Veto, Piroska Devay, Flora Szeri, Bastian Hengerer, Csaba Barta, Tamas Aranyi
    Dyskinesias are characterized by abnormal repetitive involuntary movements due to dysfunctional neuronal activity. Although levodopa-induced dyskinesia, characterized by tic-like abnormal involuntary movements, has no clinical treatment for Parkinson’s disease patients, animal studies indicate that Riluzole, w...

    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 aging DNA methylome reveals environment-by-aging interactions in amodel teleost
    Bertucci, E. M. et al.
    The rate at which individuals age underlies variation in life history and attendant health and disease trajectories. Age specific patterning of the DNA methylome (“epigenetic aging”) is strongly correlated with chronological age in humans and can be modeled to produce epigenetic age predictors. However, ...

    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...

    Perturbed DNA methylation by sustained overexpression of Gadd45b induces chromatin disorganization, DNA strand breaks and dopaminergic neurondeath in mice
    Ravel-Godreuil, C. et al.
    Heterochromatin disorganization is a key hallmark of aging and DNA methylation state is currently the main molecular predictor of chronological age. The most frequent neurodegenerative diseases like Parkinson disease and Alzheimer’s disease are age-related but how the aging process and chromatin alterations ar...

    The Identification of a Novel Fucosidosis-Associated Mutation: A Case of a5-Year-Old Polish Girl with Two Additional Rare Chromosomal Aberrations andAffected DNA Methylation Patterns.
    Domin A. et al.
    Fucosidosis is a rare neurodegenerative autosomal recessive disorder, which manifests as progressive neurological and psychomotor deterioration, growth retardation, skin and skeletal abnormalities, intellectual disability and coarsening of facial features. It is caused by biallelic mutations in encoding the α-...

    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 ...

    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...

    DNA CpG methylation in sequential glioblastoma specimens.
    Kraboth, Z and Galik, B and Tompa, M and Kajtar, B and Urban, P andGyenesei, A and Miseta, A and Kalman, B
    PURPOSE: Glioblastoma is the most aggressive form of brain tumors. A better understanding of the molecular mechanisms leading to its evolution is essential for the development of treatments more effective than the available modalities. Here, we aim to identify molecular drivers of glioblastoma development and recurr...

    Early Life Exposure to Environmentally Relevant Levels of Endocrine Disruptors Drive Multigenerational and Transgenerational Epigenetic Changes in a Fish Model
    Major Kaley M., DeCourten Bethany M., Li Jie, Britton Monica, Settles Matthew L., Mehinto Alvine C., Connon Richard E., Brander Susanne M.
    The inland silverside, Menidia beryllina, is a euryhaline fish and a model organism in ecotoxicology. We previously showed that exposure to picomolar (ng/L) levels of endocrine disrupting chemicals (EDCs) can cause a variety of effects in M. beryllina, from changes in gene expression to phenotypic alterations. Here ...

    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...

    Dnmt3a and Dnmt3b-Decommissioned Fetal Enhancers are Linked to Kidney Disease
    Guan Y, Liu H, Ma Z, Li SY, Park J, Sheng X, Susztak K
    BACKGROUND: Cytosine methylation is an epigenetic mark that dictates cell fate and response to stimuli. The timing and establishment of methylation logic during kidney development remains unknown. DNA methyltransferase 3a and 3b are the enzymes capable of establishing methylation. METHODS: We generated mice with gen...

    Rheumatoid Arthritis Patients, Both Newly Diagnosed and Methotrexate Treated, Show More DNA Methylation Differences in CD4+ Memory Than in CD4+ Naïve T Cells
    Guderud Kari, Sunde Line H., Flåm Siri T., Mæhlen Marthe T., Mjaavatten Maria D., Lillegraven Siri, Aga Anna-Birgitte, Evenrød Ida M., Norli Ellen S., Andreassen Bettina K., Franzenburg Sören, Franke Andre, Haavardsholm Espen A., Rayner Simon, Gervin Kris
    Rheumatoid arthritis (RA) is a chronic autoimmune disease that causes pain and swelling of multiple joints in the body. The underlying disease mechanisms are believed to involve a complex interplay between common genetic and environmental factors. The heritability of RA has been estimated to be ~50% for anti-citrull...

    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...

    Mitochondrial stress triggers a pro-survival response through epigenetic modifications of nuclear DNA.
    Mayorga L, Salassa BN, Marzese DM, Loos MA, Eiroa HD, Lubieniecki F, García Samartino C, Romano PS, Roqué M
    Mitochondrial dysfunction represents an important cellular stressor and when intense and persistent cells must unleash an adaptive response to prevent their extinction. Furthermore, mitochondria can induce nuclear transcriptional changes and DNA methylation can modulate cellular responses to stress. We hypothesized ...

    Differential DNA methylation of potassium channel KCa3.1 and immune signalling pathways is associated with infant immune responses following BCG vaccination.
    Hasso-Agopsowicz M, Scriba TJ, Hanekom WA, Dockrell HM, Smith SG
    Bacillus Calmette-Guérin (BCG) is the only licensed vaccine for tuberculosis (TB) and induces highly variable protection against pulmonary disease in different countries. We hypothesised that DNA methylation is one of the molecular mechanisms driving variability in BCG-induced immune responses. DNA methylatio...

    In Situ Fixation Redefines Quiescence and Early Activation of Skeletal Muscle Stem Cells
    Machado L. et al.
    Summary State of the art techniques have been developed to isolate and analyze cells from various tissues, aiming to capture their in vivo state. However, the majority of cell isolation protocols involve lengthy mechanical and enzymatic dissociation steps followed by flow cytometry, exposing cells to stres...

    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...

    DNA methylation and alcohol use disorders: Progress and challenges
    Zhang H. and Gelernter J.
    Background and Objectives Risk for alcohol use disorders (AUDs) is influenced by gene–environment interactions. Environmental factors can affect gene expression through epigenetic mechanisms such as DNA methylation. This review outlines the findings regarding the association of DNA methylation and AUDs. ...

    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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