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Methylation Analysis by Sequencing
Many scientists see mapping of the human methylome as a logical follow-up to the Human Genome Project. Methylation status of DNA varies with age, tissue type, and environmental interactions. A number of diseases such as cancer are marked by altered methylation.
Because the methylome is much more variable than the genome, methylation mapping is inherently more complex than genome sequencing. We offer several strategies to help streamline your research studies.
Figure 1. DNA methylation. A methyl group addition to the cytosine carbon 5 in cytosine-phosphate-guanine (CpG) and other nucleotide sequences inhibits the binding of transcription factors to promoters.
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Comprehensive, genome-wide mapping of methylation sites
Next-generation sequencing platforms enable high-throughput sequencing, mapping, and counting of short DNA reads (tags). In combination with traditional methylation profiling strategies (bisulfite conversion, digestion with methylation-sensitive restriction enzymes, and antibody- or 5-methylcytosine binding protein–based purification), NGS allows for comprehensive, genome-wide mapping of methylation sites.
Targeted methylation with next-generation sequencing
For targeted analysis of methylation for specific regions, the gold-standard technique is bisulfite conversion and DNA sequencing.
Methylation analysis by next-generation sequencing
Three genome-wide methylation profiling strategies are available for the Ion GeneStudio S5 series.
- Bisulfite conversion
Chemical modification of all unmethylated cytosines in the genome - Enzymatic genomic partition
Separate the genome into methylated and unmethylated compartments with methylation-sensitive restriction enzymes - Enrichment of methylated DNA
Affinity purification of methylated genomic DNA fragments
Methylation analysis—sample preparation for NGS
Epigenomic information is important in understanding how the function of the genetic sequence is implemented and regulated.
MSMSA (methylation sensitive mobility shift assay)
MSMSA is a DNA fragment analysis application used to assess the degree of methylation present in a given amplicon.
High accuracy, short time-to-answer, and low input requirements
With Ion Torrent focused next-generation sequencing (NGS) technology, get the speed, scalability and precision you need to spend more time finding answers and less time looking for them. Further your epigenetics research with our comprehensive NGS solution, which enables a complete, targeted 3-day* workflow for quantitative methylation analysis across multiple loci when combined with the bisulfite conversion method.
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For Research Use Only. Not for use in diagnostic procedures.