Methylated DNA Immunoprecipitation (MeDIP) is considered a DNA extraction technique. An antibody-immunoprecipitation reaction occurs between the DNA of interest and antibodies specific to methylated cytosines (5mC) to isolate methylated DNA fragments.
MeDIP Steps:17
- Purification of DNA from sample
- Sonication of DNA into fragments ranging in size between 300–1000 bp
- Denaturation
- Incubation of DNA fragments with anti-5mC (affinity to methylated DNA fragments)
- Incubation of magnetic beads (secondary antibody with affinity to primary antibody)
- DNA bound to the antibody complex (methylated DNA) is separated from the rest of the DNA
- DNA elution and purification
- Immunoprecipitation and washes
- Locus-specific MeDIP-PCR or other Genome-wide array analysis
Methylated DNA fragments isolated by MeDIP can be subsequently used in library preparation methods for sequencing-based analysis (MeDIP-seq), or DNA microarrays (MeDIP-chip).
17. Karpova, Nina and Umemori, Juzoh. "Protocol for Methylated DNA Immunoprecipitation (MeDIP) Analysis." Epigenetic Methods in Neuroscience Research, 2016, Volume 105, ISBN: 978-1-4939-2753-1.
