DNA Methylation Analysis Service

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.

Fig 1. DNA methylation (5mC)

Creative BioMart offers a variety of DNA methylation analysis services based on bisulfite conversion, treatment of methylation sensitive restriction enzyme and methylated DNA enrichment strategies. Different techniques provide information on DNA methylation at different levels, spanning from single site methylation in specific genes to genome-wide methylation content.

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).

MeDIP-based Service
Methylated DNA immunoprecipitation (MeDIP) is an antibody-based, large-scale technique used to enrich and capture methylated DNA fragments for gene-specific DNA methylation studies on a genome wide scale. Following the MeDIP, DNA methylation can be analyzed using multiple downstream applications, including MeDIP-Seq, MeDIP-PCR and MeDIP-chip.

Methyl-CpG-Binding Domain Sequencing
Methyl-CpG-binding domain sequencing (MBD-Seq) utilizes the methyl binding domain of methyl binding protein MBD2 to capture methylated DNA fragments to perform next-generation sequencing. Affinity-based capture approach provides a cost-effective technique because the goal of this approach is only to sequence the methylated portion of the genome.

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.

Global DNA 5mC Quantification by LC-MS/MS
Liquid chromatography tandem mass spectrometry/mass spectrometry (LC-MS/MS) provides absolute measures of 5mC content in DNA samples that are independent of DNA quality. It’s highly accurate and sensitive and is regarded as the ‘gold standard’ for global DNA methylation measurement.

Global DNA 5mC ELISA Quantification
ELISA-based assays are typically prone to high variability; thus, they are only suitable for the rough estimation of DNA methylation. Still, they are quick and easy to perform methods that serve well for the identification of large changes in global DNA methylation.

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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For Research Use Only. Not for use in diagnostic or therapeutic procedures.


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