Diagenode

WGBS service (Whole Genome Bisulfite Sequencing)

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Catalog Number
Format
G02040000

Whole-genome bisulfite sequencing (WGBS) is the most comprehensive next generation sequencing  for DNA methylation profiling, allowing single-base resolution of 5-mC within the whole genome. By comparing treated and untreated sequences, the location of the methylated cytosines are determined.

Highest coverage for DNA methylation studies

  • Genome-wide screening for DNA methylation and single-base resolution
  • High coverage for 5-mC in low density and repeat regions
  • Dedicated scientist drives each project with high touch communication

  • Comprehensive service – Bisulfite conversion, library preparation, sequencing, and analysis

  • Description

    WGBS services include:

    QC of the genomic DNA

    • Measurement of DNA concentration
    • Assessment of the DNA quality

    Preparation of WGBS libraries*

    • gDNA shearing on Bioruptor® Pico
    • Library preparation (end repair, adenylation, methylated adaptor ligation)  
    • Size selection  
    • Bisulfite conversion
    • Library amplification  QC of the WGBS libraries (DNA concentration, analysis of the profile)

    Deep sequencing

    • Samples are sent for sequencing on an Illumina platform  
    • Single-read or paired-end  
    • 50bp to 150bp read length  
    • Coverage to be defined

    *Full protocol details can be viewed in the manual from the  Premium Whole Genome Bisulfite Sequencing (WGBS) Kit

  • Bioinformatic analysis

    Standard analysis

    Methylation calling:

    Alignment of bisulfite sequencing data to reference genome and determination of methylation status of CpG nucleotides.

    Additional analysis on request:  

    Differential methylation analysis:

    Comparison of methylation status of CpG nucleotides between sample groups.

    Annotation with genomic regions:

    Annotation of differentially methylated CpGs or of DMRs with genomic regions such as introns, exons, enhancers (when available), promoters, intergenic regions.  

    Gene ontology terms analysis:

    Enrichment analysis on gene sets. Gene Ontology terms that are overrepresented in differentially bound regions may indicate the underlying biological processes involved.  

    Pathway analysis:

    Identify biochemical pathways in which genes associated with differentially methylated regions (or individual differentially methylated CpGs) may be overrepresented.  

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