DNA methylation is an epigenetic modification related to gene regulation, which is essential for biological health and disease. DNA methylation is a biochemical process in which the DNA methyltransferases (DNMTs) catalyze the methylation of DNA bases (usually cytosines) at the 5-carbon position. The family of DNMTs include DNMT1, DNMT2, DNMT3A, DNMT3B, and DNMT3L. In mammals, DNA methylation occurs on cytosines in any context of the genome, and the majority of DNA methylation occurs in a CpG dinucleotide context in somatic cells. DNA methylation plays a very important role in key processes such as genomic imprinting, X-chromosome inactivation, and inhibition of transcription and translocation of repetitive elements. When DNA methylation is dysregulated, it may lead to diseases such as cancer.
Several methods have been developed to determine the methylation status of DNA samples. Bisulfite sequencing is a commonly used technique to detect methylation using the principle that bisulfite treatment mediates the deamination of cytosine to uracil, but that methylated cytosine residues are resistant to this conversion. Methylated cytosines can be detected by comparison of sequencing results between untreated DNA samples and the bisulfite treated samples. Bisulfite conversion is the first step in many methods for analyzing DNA methylation of specific regions of interest. Combined with next-generation sequencing (NGS) technology, bisulfite sequencing can be extended to DNA methylation analysis across an entire genome. The technique of high-performance liquid chromatography-ultraviolet (HPLC-UV) is an assay for quantifying the amount of deoxycytidine and methylated cytosines present in a hydrolysed DNA sample. Liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) is an alternative high-sensitivity approach to HPLC-UV, which requires much smaller quantities of the hydrolysed DNA sample. Enzyme-linked immunosorbent assay (ELISA) can be used for the quick assessment of DNA methylation status, but only suitable for the rough estimation of DNA methylation. Assessing the methylation level of long dispersed nuclear element-1 (LINE-1) retrotransposons can well reflect the changes in DNA methylation. Other methods for detecting DNA methylation are methyl-sensitive cut counting using methylation-sensitive restriction endonucleases, PCR-based amplification fragment length polymorphism (AFLP), restriction fragment length polymorphism (RFLP), the luminometric methylation assay (LUMA) technique, etc.
Amerigo Scientific offers a variety of reagents and kits for detecting, quantifying, or purifying methylated DNA. These include methylation controls, validated primer sets for methylation studies, kits for bisulfite conversion treatment, and kits for enrichment of methylated DNA using chromatin immunoprecipitation (ChIP). We also provide methylation assay panels designed for the quantitative methylation profiling of functionally related genes, such as tumor suppressor genes, pluripotency-associated genes, and more.
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