DNA methylation is a major epigenetic modification of genomic DNA. In recent years, a large number of studies have shown that DNA methylation plays a crucial role in maintaining normal cell function, transmitting genomic genetic imprints, embryonic development, and human tumorigenesis, whereby methylation studies become epigenetic, and even Hotspots in life science research.
So how do you detect the methylation level or methylation site of a single gene or whole genome?
The most commonly used detection protocol in current research is the use of sulfite to convert unmethylated cytosine (C) in DNA to uracil (U), while methylated cytosine remains unchanged through design specificity. The methylation primers are subjected to methylation-specific PCR or sulfite sequencing to achieve experimental purposes. So how to ensure that 100% of the unmethylated cytosine in the DNA has been transformed, preventing DNA degradation during the transformation process, and the effective recovery of DNA after transformation has become the key!
Many researchers will enrich methylated DNA before bisulfite conversion. One can reduce the background, and the second can also save costs. There are currently two basic methods, one is the use of 5'-methylcytosine antibody to capture methylated DNA, namely methyl-DNA immunoprecipitation (MeDIP); the other is the use of the methyl-CpG binding domain (MBD) protein to get the same result.
Most of the products currently on the market use the latter method. The difference between the two methods is that MeDIP specifically recognizes 5'-methylcytosine in the context of single-stranded DNA, while the latter prefers double-stranded DNA. Another advantage of MBD is that when the immunoprecipitated DNA is eluted using proteinase K, the methylated DNA in MBD2 can be gradually released according to the salt gradient. This way, you can distinguish between different levels of methylated DNA.
As a professional solution provider for epigenetics, Aimei is pleased to recommend Epigentek's BisulFlash DNA Bisulfite Conversion Mag-96 Kit. As the newly developed magnetic bead method - DNA sulfite conversion kit, this kit provides optimized buffer formulations and reagents for ultra-fast bisulfite conversion of DNA samples. The transformed DNA is bound by magnetic beads, washed, eluted, and purified against column purification and agarose beads beads, effectively reducing the interference caused by non-specific adsorption!
This kit is a 96-well plate-based DNA sulfite conversion kit for high-throughput studies with the following benefits:
- Fast: 96 samples are operated at the same time, only 1h20min, and only 50s per sample.
- Convenient: Provides a mixture of conversion fluids that can be used directly in liquid form without complex configuration.
- Efficient: Complete CT transformation while DNA denaturation.
- Efficiency: The conversion efficiency has reached 99.99% of the other similar kits on the market, and the error conversion rate is as low as 0.1%. The experimental results are more reliable.
- Flexible: You can design a single experiment or choose a high-throughput study.
*This kit needs to be equipped with a 96-well magnetic frame.
(EpiMag HT (96-Well) Magnetic SeparatorCat#Q10002-1)
Figure.2 Schematic diagram of kit operation flow Figure.3 Effect of DNA after transformation, qPCR verification
Epigentek Group is the world's leading developer and supplier of innovative technology and epigenetic research products. The company has developed a comprehensive product portfolio of more than 400 patented products to provide comprehensive system solutions for epigenetic research and new drug development. As the general agent of Epigentek Group's China region, Amy Jie introduced it to China as its regional agent in China and will provide customers with complete epigenetic solutions, including analytical kits for antibodies, antibodies and related Products such as inhibitors/antagonists. At the same time, as a Chinese agent or regional agent of many international famous brands, Aimei can provide you with a series of laboratory solutions. If you are interested in the above products, please call the free customer line 400-6800-868 to contact Aimei Technology Co., Ltd. for product information and complete experimental solutions. Http://
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