This video demonstrates the protocol for methylated DNA immunoprecipitation (MeDIP). MeDIP is a two day procedure that selectively extracts methylated DNA fragments from a genomic DNA sample using antibodies with specificity for 5 -methylcytosine (anti-5 mC).
There is a growing awareness of the significant role DNA methylation plays in disease, therefore the development of assays to measure this modification are becoming increasingly important 3, 12, 13. The MeDIP technique is an amenable tool for screening at both the whole-genome and locus-specific level 6, 7. This technique provides a rapid view of DNA methylation levels using limited amounts of starting DNA and allows for easy comparisons between different sources. Downstream applications using th…
We wish to thank members of the Brown and Lam labs for their participation in critiquing this video and article. This work was supported by funds from the Canadian Institutes for Health Research and the Michael Smith Foundation for Health Research.
Material Name | Type | Company | Catalogue Number | Comment |
---|---|---|---|---|
Biorupter sonicator | Tool | Diagenode | UCD-200 TM | |
1.7ml SafeSeal Microcentrifuge Tubes | Other | Sorenson BioScience | 11510 | |
ND 3300 Spectrophotometer | Tool | NanoDrop | ||
Primary Antibody: Anti-5′-methylcytosine mouse mAb | Reagent | CalBiochem | 162 33 D3 | |
Secondary Antibody: Dynabeads M-280 Sheep anti-mouse IgG | Reagent | Invitrogen | 112-01D | |
Magnetic Tube Rack | Tool | Invitrogen | CS15000 | |
Mini LabRoller | Tool | Labnet International | H5500 | |
IP Buffer | 10 mM NaPO4 pH 7.0, 140 mM NaCl, 0.05% Triton X-100. Stored at room temperature | |||
Digestion Buffer | 10 mM Tris pH8.0, 100 mM EDTA, 0.5% SDS, 50 mM NaCl |