Aging is a complex biological process that involves a myriad of genetic and environmental factors, leading to alterations in DNA methylation patterns. Methylation-specific PCR (MSP) is a specialized technique that enables the detection and quantification of DNA methylation patterns in CpG islands, providing valuable insights into the epigenetic changes associated with aging. For performing MSP, DNA is modified by PCR performed with two primer pairs, which are detectable methylated and unmethylated DNA, respectively. MSP is a rapid measure for assession of the methylation status in CpG island.
Fig. 1 Principles of bisulfite treatment and MSP. (Muraki Y, et al. 2009)
CD BioSciences is dedicated to advancing the understanding of aging through reliable methylation-specific PCR services. From optimized sample processing to advanced data analysis, we offer unparalleled expertise and technologies.
Sample Requirements
At CD BioSciences, our MSP services require high-quality DNA extracted from diverse sources at different ages, including tissues or cell lines. Accurate and reliable results in MSP necessitate careful consideration of sample quality and quantity. Additionally, we emphasize the significance of optimized sample collection and preservation methodologies for aging-related studies.
Sample types |
Minimum input quantity |
DNA |
200 ng |
Cells |
0.2-1 M |
Tissues |
5 mg |
Workflow of Methylation-Specific PCR in Aging Research
Our workflow of MSP encompasses several crucial steps, each contributing to the specificity of the methylation analysis.
- DNA extraction. We begin with DNA extraction from the sample of aging or aging-related diseases. High-quality and intact DNA is essential to ensure reliable methylation analysis.
- Bisulfite conversion. We perform bisulfite treatment for extracted DNA, during which unmethylated cytosines are converted to uracil, while methylated cytosines remain unaltered. Our chemical conversion services are crucial for distinguishing between methylated and unmethylated DNA sequences.
- PCR amplification. Following bisulfite conversion, we employ PCR amplification using methylation-specific primers. These primers are designed to specifically target regions of interest that contain methylated DNA sequences. The amplification step allows for the selective amplification of methylated DNA, enabling the detection and quantification of DNA methylation at specific loci.
- Analysis of amplified products. We then analyze the amplified products from the PCR using gel electrophoresis or quantitative techniques. Gel electrophoresis can visualize the presence or absence of methylated DNA based on the size or intensity of the DNA bands. Quantitative methods, such as real-time PCR, provide quantification of DNA methylation levels at specific CpG sites.
Our Service Features
- Customized primer design. We possess the ability to design and validate methylation-specific primers tailored to specific genomic regions of interest. Our personalized approach allows for precise investigation of age-related methylation patterns.
- High-throughput analysis. We have the capacity for high-throughput analysis of multiple CpG sites associated with aging-related genes. We can process a large number of samples efficiently.
- Quality control. We perform the implementation of rigorous quality control at every stage of the workflow, including sample processing, amplification, and data analysis, to ensure the reliability and reproducibility of results.
CD BioSciences' MSP services exhibit a spectrum of features tailored to meet the intricate requirements of aging-related studies. Our advanced technology and expertise enable comprehensive DNA methylation profiling, encompassing high-throughput analysis of multiple CpG sites associated with aging-related genes. If you are interested in our services, please feel free to contact us or make an online inquiry.
Reference
- Muraki Y, et al. Epigenetic DNA hypermethylation: clinical applications in endometrial cancer (Review). Oncol Rep, 2009, 22 (5), 967-72.
All of our services and products are intended for preclinical research use only and cannot be used to diagnose, treat or manage patients.