Targeted Bisulfite Sequencing is an NGS method used to evaluate site-specific DNA methylation changes. The process is similar to pyrosequencing but offers a much higher throughput overall. Our next-generation sequencing platforms can deliver a great amount of useful DNA methylation information with publication-ready data parsed by our expert bioinformatics scientists. The methylation analysis at single-base resolution of individual cytosine in DNA is facilitated by bisulfite treatment of DNA followed by PCR amplification of targeted region, library construction, and sequencing of the amplicon regions. Specific primers are designed for the region of interest and cytosine methylation changes are evaluated within that region. Each DNA methylation site of interest receives a high-sequencing depth of coverage for accurate, quantitative and single-base resolution data output.
EpiGentek’s Targeted Methyl-Seq platform yields reliable information on the methylation states of individual cytosines within the targeted region by effectively and efficiently preparing converted DNA for use in next-generation sequencing techniques.
Example Data
*These are examples of data that may be provided. Please consult your service representative regarding the exact data you will receive.
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- Histogram CpG site coverage of sample set using EpiGentek’s DNA Methylation Analysis services.
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- Differential methylation annotation provides a bar chart of differentially methylated sites overlapping with genomic features (promoters, exons, introns, CpG islands, and CpG shores
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- Histogram distance TSS provides distances of differentially methylated sites to the nearest transcription start site.
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- A histogram of average read depth coverage for EpiGentek’s DNA Methylation Analysis services.
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- Methylation correlation provides sample correction shown in scatter plots (lower left) and correlation coefficients (upper right).
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- The Methylation Heatmap offers statistical grouping and visualization of samples based on DNA methylation levels of CpG sites. Green = 100% methylated; Red = 0% methylated.
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- Histogram % CpG methylation provides CpG methylation distribution within sample. Normally there should be two peaks at both ends.
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- Read alignment views of DNA Methylation Analysis data at 19 kb and base pair resolution in chromosome 7 using Integrative Genomics Viewer (IGV). The data shows CpG methylation differences for treated and control samples. Control samples are unmethylated (blue) while treated samples are methylated (red).
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- The Methylation Heatmap offers statistical grouping and visualization of samples based on DNA methylation levels of CpG sites. Green = 100% methylated; Red = 0% methylated.
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- Bismark Mapping aligns bisulfite treated sequences to a genome of interest and will output comprehensive methylation data that can be visualized in a genome viewer or used for further analysis.
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- Results Tables provide information about individual CpG sites and the frequency of methylation in addition to differential methylation analysis results. For each particular base, the methylation difference value is given along with methylation status as hyper- or hypo- methylated, as well as the chromosomal position and pvalue.
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- The Methylation Bedgraph allows quick visual comparison of DNA methylation for each chromosome in the genome.
Sample Requirements
We currently accept DNA, cell, and tissue samples for our Targeted Bisulfite Sequencing Methyl-Seq service. Proper sample preparation along with the appropriate quality control methods on your end will allow the greatest chance for assay success. Please follow all guidelines for sample requirements, packaging and labeling, and shipping which are outlined on the
Submission Instructionspage.
Talk to a Scientist
Want more information on our Targeted Bisulfite Sequencing Methyl-Seq service or interested in customizing your research project? We’re here to help! Get in touch with one of our highly knowledgeable epigenetic services experts by calling toll-free at 1-877-374-4368 or emailing services @ epigentek.com.