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DNA methylation can be detected by the following assays currently used in scientific research: [101] Mass spectrometry is a very sensitive and reliable analytical method to detect DNA methylation. MS, in general, is however not informative about the sequence context of the methylation, thus limited in studying the function of this DNA modification.
Bisulfite sequencing applies routine sequencing methods on bisulfite-treated genomic DNA to determine methylation status at CpG dinucleotides. Other non-sequencing strategies are also employed to interrogate the methylation at specific loci or at a genome-wide level. All strategies assume that bisulfite-induced conversion of unmethylated ...
The first epigenetic modification to be characterized in depth was DNA methylation. As its name implies, DNA methylation is the process by which a methyl group is added to DNA. The enzymes responsible for catalyzing this reaction are the DNA methyltransferases (DNMTs). While DNA methylation is stable and heritable, it can be reversed by an ...
This method is an extension of bisulfite sequencing, which is the gold standard for determining DNA methylation. [2] NOMe-seq relies on the methyltransferase M.CviPl, which methylates cytosines in GpC dinucleotides unbound by nucleosomes or other proteins , creating a nucleosome footprint.
The first few steps of COBRA, and the molecular changes caused by each step to methylated and unmethylated CpG sites. Combined Bisulfite Restriction Analysis (or COBRA) is a molecular biology technique that allows for the sensitive quantification of DNA methylation levels at a specific genomic locus on a DNA sequence in a small sample of genomic DNA. [1]
These methods were limited by the inability to amplify methylated DNA via polymerase chain reaction in vitro due to loss of methylation status. [6] As a result, much of these early methods relied on detecting and analyzing naturally-manifested methylated cytosines in vivo rather than chemically methylated cytosines.
The modification may also occur at other sites, [4] but methylation at either of these sites can repress gene expression by either interfering with the binding of transcription factors or modifying chromatin structure to a repressive state. [5] Disease condition studies have largely fueled the effort in understanding the role of DNA methylation.
Bisulfite methods, such as used by RRBS, were also found more accurate than enrichment based, such as MeDip-Seq. [7] The data obtained on RRBS and the Illumina Infinium methylation are highly comparable, with a Pearson correlation of 0.92. [7] The data for both platforms are also directly comparable as both use an absolute measurement of DNA.