Methods to analyze DNA methylation, DNA demethylation, and their functional effects are critical to epigenetics researchers. Methylation profiles of epigenome are used for disease identification and for research and therapeutic development. Analysis can be gene-specific or global depending on downstream applications (1).
Downstream methylation detection methods include end-point PCR, real-time PCR, primer extension, single-stranded conformational polymorphism assays, blotting, microarray analysis and sequencing. Choosing which method(s) to use largely depends on the experimental sample size and the goals of the experiments.
- Tao Zuo et al. 2009, Epigenomics 1(2) 331-345. PMID: 20526417
- Do I need to do extra purification steps for a mammalian DNA prep in order to use it in a glucosylation reaction?
- How much control DNA should I use in my qPCR?
- Does the T4-BGT show any site preference?
- How do I know if the glucosylation of 5-hmC in my DNA is complete?
- What is the shelf life of the reagents supplied?
- What products does NEB offer for the study of Epigenetics?
Epigenetics - Expanding on Genomic Foundations
- Epigenetics Brochure
- EpiMark® Methylated DNA Enrichment Kit Troubleshooting Guide
- Woodcock, C.B.l, Yu, D., Hajian, T., Li, J., Huang, Y., Dai, N., Corrêa, I.R., Jr., Wu, T., Vedadi, M., Zhang, X., Cheng, X (2019) Human MettL3 MettL14 complex is a sequence specific DNA adenine methyltransferase active on single strand and unpaired DNA in vitro Cell Discov ; 5:63, PubMedID: 31885874, DOI: 10.1038/s41421-019-0136-4
- Pais, J.E., Dai, N., Tamanaha, E., Vaisvila, R., Fomenkov, A.I., Bitinaite, J., Sun, Z., Guan, S., Correa, I.R. Jr., Noren, C.J., Cheng, X., Roberts, R.J., Zheng, Y. and Saleh, L. (2015) Biochemical characterization of a Naegleria TET-like oxygenase and its application in single molecule sequencing of 5-methylcytosine Proc Natl Acad Sci U S A; 112(14), 4315-4321. PubMedID: 25831492
- Pirone-Davies, C., Hoffmann, M., Roberts, R.J., Muruvanda, T., Timme, R.E., Strain, E., Luo, Y., Payne, J., Luong, K., Song, Y., Tsai, Y.C., Boitano, M., Clark, T.A., Korlach, J., Evans, P.S., Allard, M.W. (2015) Genome-Wide Methylation Patterns in Salmonella enterica Subsp. enterica Serovars PLoS One; 10(4), e0123639. PubMedID: 25860355
- Hughes J.R., Roberts N., McGowan S., Hay D., Giannoulatou E., Lynch M., De Gobbi M., Taylor S., Gibbons R., Higgs D.R. (2014) Analysis of hundreds of cis-regulatory landscapes at high resolution in a single, high-throughput experiment Nat Genet; 46 (2), 205-212. PubMedID: 24413732, DOI: doi:10.1038/ng.2871
- Sexton T, Kurukuti S, Mitchell JA, Umlauf D, Nagano T, Fraser P (2012) Sensitive detection of chromatin coassociations using enhanced chromosome conformation capture on chip Nat Protoc; 7(7), 1335-50. PubMedID: 22722369, DOI: 10.1038/nprot.2012.071
- Bisulfite conversion, which provides single nucleotide, high resolution data for whole genomes or single locus methylation studies.
- Differential enzymatic cleavage, which uses methylation, or dually 5-mC and 5-hmC dependent enzymes, paired with non-sensitive isoschizomers to asses DNA methylation or DNA hydroxymethylation.
- High-Performance Liquid Chromatography (HPLC)
- Matrix-Assisted Laser Desorption/Ionization-Time of Flight Mass Spectrometry(MALDI-TOF MS)
- Methylated DNA-ELISA
- Affinity-based methods, which use methylated DNA binding proteins or antibodies for enrichment.
- Enzymatic DNA methyltransferase assay
Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email firstname.lastname@example.org.
This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
If all cells are created from the same genetic material, why are there so many different cell types? Listen to Sriharsa Pradhan, Senior Scientist, RNA Biology at NEB, as he describes how DNA is methylated and how this affects the path of reading the DNA code the same way an obstruction would derail a train off its tracks.