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Premium RRBS kit V2
RRBS for low DNA amounts and accurate analysis

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Catalog Number
Format
C02030036
24 rxns

Reduced Representation Bisulfite Sequencing (RRBS) offers a cost-effective, focused 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
  • 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.

Premium RRBS kit V2 includes all reagents necessary for the library preparation. Specific adapters were designed and validated to fit the Premium technology and are available separately:

For RRBS UDI-UMI library construction:

How Do UMIs Work?

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

Figure 1. Premium RRBS V2 construct. Integrated UMIs enable removal of PCR duplicates during the data analysis.

Accurate Coverage of CpGs

Table 1. Examples of Premium RRBS V2 sequencing data. UMI data processing enables accurate estimation of CpG counts.

  • Validation: Sequencing quality

    Figure 2. 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 and 2 (data shown for 50 ng gDNA input after trimming).

  • Validation: Focus on CpG-rich regions

    Figure 3. 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).

  • Software for Intelligent Pooling

    Diagenode's new online intelligent pooling aid (RRBS SIP) provides the optimal pool design for RRBS to meet your specific sample and analysis needs:

    • Time-saving - Avoid complex caculations
    • Highest pooling efficiency based on qPCR quantification - Bring the power of pooling to the highest
    • Powerful - Incorporates advanced aspects such as number of samples per pool required, the separation between projects, and more
    • Accurate - Identify outliers

    Get access to Diagenode's RRBS SIP - coming soon.

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    RRBS for low DNA amounts and accurate analysis (Diagenode Cat# C02030036)
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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
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