Diagenode

Targeted Bisulfite Sequencing Service

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Catalog Number
Format
G02070000

Targeted bisulfite sequencing enables DNA methylation analysis of targeted genomic regions with single nucleotide resolution. Depending on the size of the region of interest, the methodology may include a PCR-based step to amplify multiple bisulfite-converted DNA regions in a single reaction (PCR enrichment workflow) or a hybridization capture step using pre-designed oligos to isolate DNA regions that contain target CpG sites (hybridization capture workflow).

This method is useful for hypothesis testing studies of target regions of interest as well as confirmation of regions identified in genome-wide methylation analyses such as WGBS, RRBS, and MeDIP-seq.

 DNA methylation with focused content

  • DNA methylation patterning of selective regions
  • Single nucleotide resolution
  • Enhanced accuracy, sensitivity and specificity while reducing overall cost
  • Allowing deep sequencing at high coverage levels for rare variant identification
  • Suitable for epigenetic biomarker validation 

Customized end-to-end service

  • From project design to data analysis
  • A dedicated scientist driving your project with high touch communication
  • High-quality bioinformatic support to assist you with data analysis

PCR enrichment worflow

Hybridization capture worflow

See our other DNA Methylation Profiling Services

  • Service Worflow

    Targeted Bisulfite Sequencing Service will choose between two methodologies depending on the size of the region of interest:

    Workflow

    Description

    Region of interest

    PCR Enrichment

    • (gDNA shearing on Bioruptor Pico)
    • Bisulfite conversion
    • PCR primer design
    • PCR amplification optimization
    • Library preparation
    • QC of final libraries (DNA concentration, analysis of the profile)
    • Sequencing on Illumina platform
    • Several, long DNA regions
    • Total size up to 200kb

    Hybridization Capture

    • (gDNA shearing on Bioruptor Pico)
    • Bisulfite conversion
    • Library preparation
    • Capture probe design
    • Amplification optimization
    • QC of final libraries (DNA concentration, analysis of the profile)
    • Sequencing on Illumina platform
    • Large number of very long DNA regions
    • Total size between 0.2-100Mb
  • Bioinformatic Analysis

    Analysis

    Features

    Standard
    • FASTQ raw data
    • FASTQC quality control insights
    • Alignment of bisulfite sequencing data against reference genome
    • Methylation calling and extraction
    • Summary statistics
    Differential methylation analysis
    • Methylation level analysis
    • Differentially Methylated CpGs (DMCs) analysis
    • Differentially Methylated Regions (DMRs) analysis
    • Annotation of DMCs and DMRs for genomic regions (exons, introns, …) and for CpG island locations (islands, shores, shelves, ...)

  •  Documents
    Epigenomics Profiling Services FLYER
    Chromatin analysis DNA methylation services RNA-seq analysis
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  •  Publications

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