Diagenode

Premium Reduced Representation Bisulfite Sequencing (RRBS) kit

カタログ番号
フォーマット
価格
C02030032
24 rxns
$1,455.00
ブローカー情報

Reduced Representation Bisulfite Sequencing (RRBS)は単一ヌクレオチドレベルで、全ゲノムバイサルファイトシーケンシングに関連するDNAメチル化を効率的また、低コストで分析する非常に有効的な方法です。 制限MspI酵素(CCGG標的部位)およびサイズ選択を用いてゲノムを切断することで、通常、DNAメチル化マークが見出されるCpGリッチ領域(CpGアイランドを含む)を示すためにDNAが濃縮されます。 したがって、RRBSは、実際に配列決定が必要なゲノム部分を減らし、関連するCpGアイランドに焦点を当てることによって、DNAメチル化分析のコストパフォーマンスを高めます。

Diagenode, RRBS技術に関するビデオはこちら。

Reduced Representation Bisulfite Sequencing(RRBS)によるDNAメチル化解析」に関するウェビナーを見る(1時間程度)

  • Characteristics
    • Superior coverage – 4 million CpGs
    • High throughput - 96 samples processed in one experiment
    • Cost-efficient – Multiplex 6-8 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

  •  お客様の声

    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
  •  資料
    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. ...
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    Premium RRBS kit MANUAL
    Diagenode’s Premium RRBS technology Positive and negative spike-in controls are...
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    Premium RRBS spike-in controls manual 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
  •  出版物

    How to properly cite this product in your work

    Diagenode strongly recommends using this: Premium Reduced Representation Bisulfite Sequencing (RRBS) kit (Diagenode Cat# C02030032). Click here to copy to clipboard.

    Using our products in your publication? Let us know!

    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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