DNA Methylation Analysis: Role & Techniques for Detecting Epigenetic Modifications, Slides of Biology

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DNA Methylation Analysis

DNA methylation is an important epigenetic modification that regulates transcription and thus affects

gene expression. It is a popular research object in the fields of embryonic development, postnatal

development, cancer, and the influence of environmental hormones on epigenetic inheritance. In

mammals, DNA methylation is found to occur predominantly on cytosines followed by guanine residues

(CpG). This type of methylation is referred to as CpG methylation, and cytosine methylated at the fifth

carbon of the pyrimidine ring is called 5-methylcytosine (5mC). DNA methylation patterns vary across

organisms. The mammalian genome is usually highly methylated and those CpG loci that can be

methylated are not randomly distributed in the genome sequence. In the genome of healthy people,

the CpG loci in CpG islands are generally unmethylated, while the CpG loci outside the CpG islands are

usually methylated. Non-CpG methylation is prevalent in human embryonic stem cells (ES cells) and

brain tissue. Meanwhile, non-CpG methylation plays key roles in plants. Due to the influences of

environment, disease, age and gender factors, DNA methylation is in a dynamic change process, in which DNA methylation can be established and maintained by DNA methyltransferase (DNMT) and it can be demethylated by DNA demethylase. Aberrant change of DNA methylation content and patterns is a significant factor in the occurrence and development of diseases. In cancer research, abnormal change in DNA methylation considered to be one of biomarkers for early diagnosis and the promising target for developing therapeutic approaches.

DNA Methylation Analysis Services at Creative BioMart Bisulfite Conversion Service Bisulfite conversion assay is based on the chemical conversion of unmethylated cytosine to uracil, while methylated cytosine is exempt from this conversion, after which various analyses can be performed on the altered sequences to retrieve this information. Bisulfite Sequencing Service The combination of bisulfite conversion and high- throughput sequencing provides the most quantitative method for DNA methylation analysis at single-base resolution. Creative BioMart offers comprehensive bisulfite sequencing services including whole-genome bisulfite sequencing (WGBS), reduced representation bisulfite sequencing (RRBS) and targeted bisulfite sequencing.

Oxidative Bisulfite Sequencing Service Oxidative bisulfite sequencing technology first oxidizes 5hmC into 5fC, which is then converted into U by bisulfite treatment, thus eliminating the interference of 5hmC and achieving precise detection of DNA methylation. MRE-Seq Service MRE-Seq stands for methylation-sensitive restriction enzyme digestion followed by sequencing. Unmethylated CpG sites are identified at single CpG site resolution by sequencing size-selected fragments from parallel DNA digestions with the methyl-sensitive restriction enzymes (MREs).

Methylation-Specific PCR Service Methylation-specific PCR (MSP) is probably the most widely used technique for studying DNA methylation of a locus of interest. This simple, rapid, and cost-effective method can be used to specifically detect the DNA methylation status of almost any group of CpG sites within a CpG island without the requirement for methylation- sensitive restriction enzymes. MS-HRM Analysis Service Methylation-sensitive high-resolution melting (MS-HRM) is an in-tube, PCR-based method to detect methylation levels at specific loci of interest. A unique primer design facilitates a high sensitivity of the assays, enabling detection of down to 0.1-1% methylated alleles in an unmethylated background.

Bisulfite Sequencing PCR Service After DNA samples are treated with bisulfite, primers are designed for PCR amplification of the target fragment, and the PCR products are cloned and sequenced. The sequenced sequence is compared with the reference sequence to determine whether the CpG site is methylated. DNA Methylation Microarray Service Bisulfite conversion, MeDIP, MBD-based capture, and almost any other techniques can utilize microarray technology. The main advantage of microarrays in methylation studies is that they can target methylated or potentially methylated regions.

Creative BioMart has organized a team of scientists with extensive experience in DNA methylation analysis to provide a flexible, one-stop service for your DNA methylation analysis project. We can help you with experimental design, carry out your experiments and data analysis. We can provide diversity methods for you to choose as you prefer, and we can also help recommend the most appropriate solution. Moreover, we can provide a customized combination of services for comprehensive and in-depth DNA methylation analysis. Our experienced experts can help you design the optimal research solution and eliminate all the hurdles in your research. If you are interested in DNA methylation analysis, please feel free to contact us , Creative BioMart provides you with professional and thoughtful service!

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Excerpt from: https://www.creativebiomart.net/epigenetics/services/dna- methylation-analysis-service/