DNA MethylationEnlarge Image
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. Click to enlarge.

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.

Comprehensive, Genome-Wide Mapping of Methylation Sites

Next-generation sequencing platforms enable ultra–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), sequencing allows for comprehensive, genome-wide mapping of methylation sites.

And, for targeted analysis of methylation for specific regions, the gold-standard technique is bisulfite sequencing using automated capillary electrophoresis (CE) instruments.

Methylation Analysis by Sequencing Using the 5500xl Genetic Analyzer
Three genome-wide methylation profiling strategies are available for the 5500xl Genetic Analyzer.
  • Enzymatic Genomic Partition
    Separate the genome into methylated and unmethylated compartments with methylation-sensitive restriction enzymes.
  • Bisulfite Conversion
    Chemical modification of all unmethylated cytosines in the genome.
  • Enrichment of Methylated DNA
    Affinity purification of methylated genomic DNA fragments.
 Methylation Analysis Using the 3500 Genetic Analyzer
For the targeted analysis of methylation in specific regions, bisulfite sequencing using automated CE provides detailed information for each CpG across an entire amplicon. Allele-specific haplotypes are subsequently obtained via cloning.
 
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.
 

Methylation Analysis by Sequencing Sample Prep for Next Generation Sequencing
Epigenomic information is important in understanding how the function of the genetic sequence is implemented and regulated.