DNA methylation is adding methyl groups to certain bases without changing the base type or DNA sequence. It belongs to the category of epigenetics, which alters the traits of an organism mainly by affecting gene expression. Lifeasible provides DNA methylation detection solutions based on the bisulfite method, based on the principle that bisulfite can convert C bases into U bases. The methylated C-base does not react with bisulfite and remains in the C-base state throughout the treatment.

Bisulfite modification

The principle of bisulfite modification is that in the presence of HSO₃^-, single-stranded DNA can effectively convert unmethylated C to U. After two rounds of PCR amplification, m5C remains unchanged and remains C, while the site is changed to T.

• We can identify whether the site to be tested is methylated by sequencing and comparing the amplified clones. This technique makes it easy to determine the methylation position of individual single strands and their methylation status in individual genomic DNA molecules.
• On this basis, we can measure the methylation status of the corresponding sequences by combining restriction endonuclease analysis, direct DNA sequencing, methylation-sensitive single-nucleotide primer extension, methylation-specific PCR amplification, lock probe technology, and high-performance liquid chromatography.

Bisulfite-conjugated DNA direct sequencing method

We can use this method to test the methylation of specific genes, such as determining the methylation status of a gene in a certain state.

The bisulfite-bound methylation-specific PCR method

After treating the sample DNA with bisulfite, the methylated C is unchanged. At the same time, the unmethylated C is replaced by U. The methylated C is replaced by U, and the unmethylated C is replaced by U. Primer selection and design are critical, and we design two different sets of primer pairs during the PCR reaction.

• One primer pair is designed for the non-methylated DNA strand after sodium bisulfite treatment. If a fragment can be amplified with this primer pair, it means that the detection site is not methylated.
• The other pair of primers are designed for methylated DNA strands after sodium bisulfite treatment. If a fragment can be amplified with this pair of primers, it means that the test site is methylated.

We can use this method to detect the methylation level of genomic DNA.

Combined bisulfite restriction analysis (COBRA) method

We use COBRA to directly analyze the digest products, making the quantification of COBRA free of radioactive contamination more accurate and rapid. This method is relatively simple and does not require the researcher to know the sample sequence and CpG sites in advance. It also requires a small sample size and can be applied to the analysis of paraffin-embedded samples, which allows us to assist you in quantifying DNA methylation levels.

Lifeasible offers methylation detection and analysis by bisulfite modification, bisulfite-conjugated DNA direct sequencing, bisulfite-conjugated methylation-specific PCR, and COBRA. Please feel free to contact us for more information on our solutions or for customized solutions.

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For research use only, not intended for any clinical use.

Related Services

• Restriction Endonuclease-Based DNA Methylation Detection
• Immunoprecipitation of Methylated DNA
• SMRT Sequencing for Direct Detection of DNA Methylation
• Single Molecule Nanopore Dependent Methylation Sequencing
• RNA Methylation
• RNA-Directed DNA Methylation
• DNA Hydroxymethylation