Diagenode

Premium RRBS kit V2

目录号
格式
价格
C02030036
24 rxns
$1,480.00

(Coming soon)

Reduced Representation Bisulfite Sequencing (RRBS) offers a cost-effective, targeted 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
  • Use DNA inputs as low as 25ng
  • Obtain higher accuracy with UMI integration -- included in kit
  • 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

How Do UMIs Work?

UMIs incorporate a unique barcode onto each molecule within a given sample library. By incorporating individual barcodes on each original DNA fragment, variant alleles present in the original sample (true variants) can be distinguished from errors introduced during library preparation, target enrichment, or sequencing.

<< Image.jpg>>

Accurate Coverage of CpGs

<<Image.jpg>>

  • Validation: Sequencing quality

    Sequencing quality

    Figure 1. 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 2. 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).



    Compatible with all vertebrates

    Table 2. The kit RRBS v2 is compatible with all vertebrates. The Table 2 shows some examples of results obtained for different species.



    Superior performance

    • Lower duplicate rate
    • More CpGs detected
    • Higher % of uniquely aligned reads containing CpGs
    • Higher genome coverage (3,3%)

    Table 3. RRBS was performed using Diagenode RRBS v2 kit on HeLa cells and human blood samples (total 10 samples) as well as using competitor kit on HeLa cells, human blood and brain samples (total 10 samples). The Table 3 shows sequencing parameters for 10 samples processed with each kit.

    Figure 3. Comparison of CpG coverage between competing technologies.

  • Validation: Focus on CpG-rich regions

    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.

    NATURE METHODS

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

    Christoph Bock, Sharon SquazzoMiklos LaczikPaul DatlingerAnne-Clémence Veillard

    Nature Methods 13 (2016)

    Published online

    CHECK OUT THE NATURE METHODS PAPER

  •  文档
    Template SIP RRBS EXCEL FILE
    Template file for RRBS SIP (Software for Intelligent Pooling)
    Download
    DNA Methylation Solutions BROCHURE
    Complete solution for DNA methylation studies! Whether you are experienced or new to th...
    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 v2 manual MANUAL
    Manual Premium RRBS kit V2
    Download
    Sequence of the methylated spike-in control in FASTA format – Premium RRBS kit v2
    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 – Premium RRBS kit v2
    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 – Premium RRBS kit v2
    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
  •  出版物

    How to properly cite this product in your work

    Diagenode strongly recommends using this: Premium RRBS kit V2 (Diagenode Cat# C02030036). Click here to copy to clipboard.

    Using our products in your publication? Let us know!

    When left does not seem right: epigenetic and bioelectric differencesbetween left- and right-sided breast cancer.
    Sofía, Masuelli and Sebastián, Real and Emanuel, Campoy andBranham, María Teresita and Marzese, Diego Matías andMatthew, Salomon and De Blas, Gerardo and Rodolfo, Arias andMichael, Levin and María, Roqué
    BACKGROUND: During embryogenesis lateral symmetry is broken, giving rise to Left/Right (L/R) breast tissues with distinct identity. L/R-sided breast tumors exhibit consistently-biased incidence, gene expression, and DNA methylation. We postulate that a differential L/R tumor-microenvironment crosstalk generates diff...

    Comparative Toxicogenomics of Glyphosate and Roundup Herbicidesby Mammalian Stem Cell-Based Genotoxicity Assays andMolecular Profiling in Sprague-Dawley Rats.
    Mesnage R. et al.
    Whether glyphosate-based herbicides (GBHs) are more potent than glyphosate alone at activating cellular mechanisms, which drive carcinogenesis remain controversial. As GBHs are more cytotoxic than glyphosate, we reasoned they may also be more capable of activating carcinogenic pathways. We tested this hypothesis by ...

    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...

    Adaptive Convergence of Methylomes Reveals Epigenetic Driversand Boosters of Repeated Relapses in Patient-matched ChildhoodEpendymomas and Identifies Targets for Anti-RecurrenceTherapies
    Zhao S. et al.
    Ependymoma (EPN) is the third most common brain tumor in children and frequently recurs. Here, we report an integrated longitudinal analysis of epigenetic, genetic and tumorigenic changes in 30 patient-matched repeated relapses obtained from 10 pediatric patients to understand the mechanism of recurrences. Genome-wi...

    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 andneuropathic chronic pain subtypes reveals alterations in methylation ofgenes involved in the neuro-musculoskeletal system.
    Stenz Ludwig 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 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...

    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, ...

    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...

  •  相关产品

活动

  • CNET 2022
    Gif-Sur-Yvette, France
    Jul 4-Jul 6, 2022
 查看所有活动

新闻

 查看所有新闻


The European Regional Development Fund and Wallonia are investing in your future.

Extension of industrial buildings and new laboratories.


       Site map   |   Contact us   |   Conditions of sales   |   Conditions of purchase   |   Privacy policy   |   Diagenode Diagnostics