mtDNA Methylation Microarray Service
CD Genomics is a world-leading company that can provide the best solutions for global customers in the field of microarrays. We have successfully completed much scientific research and non-clinical research projects to ensure that we provide the best products and services to customers all over the world.
In addition to the methylation of genomic DNA that occurs in the nucleus, which can be used as an epigenetic marker, the methylation of mitochondrial DNA (mtDNA) has gradually been paid more attention. In some studies on mitochondria, varying degrees of methylation modification (less than a quarter of the degree of methylation in the nucleus) have been detected. Mitochondrial function is related to the pathophysiological processes of neurological diseases and some metabolic diseases. For example, mitochondrial dysfunction plays a key role in cancer development and related treatment responses. Mitochondrial dysfunction in a wide range of human malignancies indicates that targeting mitochondria is still a promising way to develop new cancer treatment strategies. The data shows that the mitochondrial genome is largely methylated at non-CpG sites.
The Key to Mitochondrial Methylation Detection
The circular double-stranded structure of the mitochondrial genome is similar to that of the bacterial genome. Different from the nuclear genome is its complex secondary and tertiary structure, which will make the detection of mitochondrial DNA methylation more difficult. The secondary structure of mitochondrial DNA may prevent the bisulfite conversion step, leading to false-positive methylation values.
Fig 1. Map of the human mitochondrial genome. (Mechta M, et al. 2017)
The current mtDNA detection strategy is mainly the bisulfite method. However, unlike nuclear genomes, other methods can be used to unravel the secondary and tertiary structure of mtDNA. Accurate fragmentation is the key to the detection of mitochondrial DNA methylation, and the optimization of this step can ensure high quality and reliable methylation value. Avoid the low bisulfite conversion efficiency due to the circular ring structure of mtDNA, which will cause the increase of methylation measurements.
| Mitochondrial DNA Extraction |
Use the classic DNA extraction method of organelles. Collect the minimum amount of nucleic acid required for bisulfite modification (2μg concentration). |
| Linearization |
Enzyme digestion and ultrasonic treatment were used to achieve the best linearization. The application of enzyme digestion (BamHI restriction endonuclease) can break the loop of mitochondrial circular DNA, further ultrasonic treatment can make it fragment, unravel the secondary and tertiary structure, and facilitate the subsequent conversion of bisulfite. |
| Bisulfite Conversion |
Use the sulfite conversion kit. |
| MeIP |
Purification and enrichment using immunological methods. |
| Detection |
Microarray hybridization detection. |
Fig 2. Mitochondrial DNA Methylation Detection Service.
Sample Delivery Requirement
- We can detect a variety of samples such as tissues, cells, blood, etc. If you provide pure mtDNA samples, the sample concentration should not be too low;
- Samples need to be sent at low temperature;
- Multiple samples need to be distinguished, and you need to provide a sample number to distinguish.
Our Features and Benefits
- Personalized service
- Reliable, proven, and rapid processes
- Industry-leading data quality
CD Genomics provides high-quality mitochondrial DNA methylation microarray screening services. We have a well-equipped experimental platform, a large and professional team of scientists, and we are committed to cooperating with researchers from all over the world to meet the needs of our customers. Customers can contact our employees directly and provide timely feedback on their queries. If you are interested in our one-stop solution service, please contact us for more detailed information.
References
- Mechta M, Ingerslev L R, Barrès R. Methodology for accurate detection of mitochondrial DNA methylation[J]. JoVE (Journal of Visualized Experiments), 2018 (135): e57772.
- Patil V, Cuenin C, Chung F, et al. Human mitochondrial DNA is extensively methylated in a non-CpG context[J]. Nucleic acids research, 2019, 47(19): 10072-10085.
- Li W, Zhang X, Lu X, et al. 5-Hydroxymethylcytosine signatures in circulating cell-free DNA as diagnostic biomarkers for human cancers[J]. Cell research, 2017, 27(10): 1243-1257.
Our products and services are for research purposes ONLY. We do not perform any private testing services that are not for laboratory use.
