DNA Methylation Analysis via Bisulfite Sequencing
DNA methylation has been shown to be involved in a diverse array of cellular functions and diseases, including gene expression, differentiation, genomic imprinting, X chromosome inactivation, transposon silencing, regulation of chromatin structure and genome integrity, carcinogenesis, aging, and stem cell differentiation. DNA methylation patterns are dynamically regulated during development by a suite of epigenetic regulators, and the influence of the environment. In addition, DNA methylation changes are generally accepted as one of the most common molecular changes in human tumors.
The gold standard in analyzing DNA methylation is by chemical treatment of DNA using bisulfite. Bisulfite treatment converts cytosine to uracil while 5-methy cytosine is resistant to the conversion. After the treatment, genomic DNA is subject to PCR amplification, subcloning, and sequencing. The sequencing result is then compared to the original sequences, and any methylated/unmethylated cytosine is unambiguously determined. As a result, methylated Cs remains as Cs while unmethylated Cs become Ts (see figures below).
We offer comprehensive bisulfite sequencing service, including genomic DNA digestion, bisulfite conversion, target primer design, PCR amplification, subcloning, and random sequencing. For statistical representation of each cytosine in any given sample type, 15-20 individual clones need to be sequenced. Depending on your experimental needs, we can help you design best strategy to address your research questions. For more details and for price quote please send inquiry along with your target sequences to info@alphabiolab.com (email preferred).