Cytosine methylation can significantly modify temporal and spatial gene expression and chromatin remodeling. Leveraging the power of next-generation sequencing (NGS), both genome-wide analysis and targeted approaches can provide researchers with insight into methylation patterns at a single nucleotide level.
Advantages of methylation sequencing:
Many approaches leverage the high quality and sensitivity of NGS for methylation analysis. Most methods rely on bisulfite conversion of DNA to detect unmethylated cytosines. Bisulfite conversion changes unmethylated cytosines to uracil during library preparation. Converted bases are identified (after PCR) as thymine in the sequencing data, and read counts are used to determine the % methylated cytosines.
Bisulfite conversion sequencing can be done with targeted methods such as amplicon methyl-seq or target enrichment, or with whole-genome bisulfite sequencing. Additionally, alternative chemistries like OxBS and TAB-Seq can be used with NGS for identification of hydroxymethylation (5-hMc) in conjunction with methylation (5-mc) analysis.
See the Field Guide to DNA Methylation Analysis for a deep dive into methylation analysis methods.
More than 90% of the world’s sequencing data is generated using Illumina NGS technology.*
Illumina offers a fully supported workflow, from library preparation to data analysis, for methylation sequencing.
Studies of epigenetic alterations in cancer, such as aberrant methylation and altered transcription factor binding, can provide insight into important tumorigenic pathways. Learn more about analyzing epigenetic changes in cancer.
Genome-wide methylation sequencing can help researchers understand the functional mechanisms at work in complex neurological, immunological, and other diseases. Learn more about complex disease research.
The DNA methylation landscape of Chinese hamster ovary (CHO) DP-12 cells.
J Biotechnol 199 38-46 2015
Single-cell DNA methylome sequencing and bioinformatic inference of epigenomic cell-state dynamics.
Cell Rep 10 1386-97 2015
Transient acquisition of pluripotency during somatic cell transdifferentiation with iPSC reprogramming factors.
Nat Biotechnol 33 769-74 2015
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*Data calculations on file. Illumina, Inc., 2015