Premium Reduced Representation Bisulfite Sequencing (RRBS) Kit

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Catalog Number
96 rxns
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Reduced Representation Bisulfite Sequencing provides a powerful method to efficiently analyze DNA methylation at the single nucleotide level without the higher costs associated with whole genome bisulfite sequencing. By cutting the genome using the restriction MspI enzyme (CCGG target sites) followed by size selection, DNA is enriched to represent CpG-rich regions (including CpG islands), in which DNA methylation marks are typically found. Thus, RRBS provides a cost-effective method for analyzing DNA methylation by reducing the part of the genome that actually needs to be sequenced and focusing on relevant CpG islands.

  • Characteristics
    • Superior coverage – 4 million CpGs
    • High throughput - 96 samples processed ine 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


    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


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

    Nature Methods 13 (2016)

    Published online


  • Testimonials

    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

    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
  • Applications
    Bisulfite conversion
    Bisulfite modification of DNA is the most commonly used, "gold standard" method for DNA methylation studies providing single nucleotide resolution. This technology is based on the chemical conversion of unmethylated cytosine to uracil. Methylate... Read more
  • Documents
    RRBS Pooling aid EXCEL FILE
    RRBS Pooling aid
    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 MANUAL
    Diagenode’s Premium RRBS technology Positive and negative spike-in controls are...
  • Publications

    How to properly cite this product in your work

    Diagenode strongly recommends using this: Premium Reduced Representation Bisulfite Sequencing (RRBS) Kit (Diagenode Cat# C02030033). Click here to copy to clipboard.

    Using our products in your publication? Let us know!

    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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