DNA Methylation

DNA methylation is a major epigenetic modification involving the addition of a methyl group to the 5 position of cytosine by DNA methyltransferase to form 5-methylcytosine (5-mC). This epigenetic mark has the power to turn genes on or off and can be inherited through cell division. DNA methylation was the first epigenetic mark to be discovered and it plays an important role in normal human development, aging, tumorigenesis, and other genetic and epigenetic diseases.

DNA Hydroxymethylation

Similarly, DNA hydroxymethylation, caused by oxidation of 5-mC through the TET family of enzymes, was further discovered to be involved in controlling gene expression and is a key DNA demethylation process as part of the DNA methylation cycle.

Measuring 5-mC

Techniques to measure 5-mC have developed over the years and can vary from heat-based to chemical-based methods. Bisulfite conversion is a popular technique used for preparing DNA for methylation analysis on a gene-specific level. This process converts cytosine to uracil while leaving 5-methylcytosine intact to allow methylation analysis at single-nucleotide resolution. Not only is efficient conversion necessary, but recovery must be optimal due to the harsh chemical reaction which often degrades the DNA.

MethylFlash Global DNA Methylation Pre-Screening

Obtaining an overall view of a sample’s methylation status is a smart way to pre-screen your samples before pursuing downstream applications. Our MethylFlash global DNA methylation pre-screening tools can be used prior to more in depth investigation and costly sequencing. Conventional methods such as mass spectrometry or chromatography, which require expensive equipment or specialized expertise, are not necessary for this ELISA-based method. Also, unlike LUMA or LINE-1, Alu, and LTR-based assays, MethylFlash technology directly quantifies actual global DNA methylation.

Additional Methods

• Methylated DNA Immunoprecipitation (MeDIP) - an antibody-based technique used for studying gene-specific DNA methylation on a genome-wide scale
• Activity/Inhibition Assays - measure DNA methyltransferase (DNMT) and other enzymes related to the process of DNA methylation
• Methylation-specific PCR (MS-qPCR) - an amplification technique used for studying gene or sequence-specific DNA methylation.

DNA methylation is an epigenetic mechanism involving the addition of a methyl group to the 5 position of cytosine in DNA. This process can turn genes on or off and can be inherited through cell division. There are various techniques available to measure DNA methylation, including bisulfite conversion and MethylFlash global DNA methylation pre-screening. Additionally, there are several methods such as MeDIP, activity/inhibition assays, and MS-qPCR that can be used to investigate DNA methylation on a gene-specific or genome-wide scale.

Additional methods for investigating DNA methylation include an antibody-based technique known as methylated DNA immunoprecipitation (MeDIP) that is used for studying gene-specific DNA methylation on a genome-wide scale, activity/inhibition assays that measure DNA methyltransferase (DNMT) and other enzymes related to the process of DNA methylation, as well as an amplification technique involving methylation-specific PCR (MS-qPCR) that is used for studying gene or sequence-specific DNA methylation.

DNA Methylation Tools

DNA Bisulfite Conversion	DNA Methylation Quantification	Methylated DNA Immunoprecipitation (MeDIP)	DNA Methyltransferase & Demethylase Assays	Methylated DNA Amplification (MS-qPCR)
Gene-specific, base-resolution DNA methylation	Global DNA methylation	Gene-specific DNA methylation on a genome wide scale	Activity/inhibition of enzymes involved in DNA methylation cycle	Gene or sequence-specific DNA methylation