Pyrosequencing lab and DNA methylation
Pyrosequencing is a sequencing by synthesis method that provides a quantitative measure of the proportion of bases at every position within any PCR-amplified sequence. We exploit this property of Pyrosequencing to obtain highly quantitative measures of DNA methylation in gene sequences as well as repetitive elements that allow for discriminating differences lower than 1% methylation. Our laboratory is equipped with two Pyrosequencing PSQ™HS 96 (Pyrosequencing Inc., Westborough, MA). Seven Biorad PCR units are used for pre-Pyrosequencing amplification.
Pyrosequencing has been successfully applied to investigate DNA methylation in population environmentally or occupationally exposed to several toxicants, such as Particulate Matter (PM), Heavy Metals, Polycyclic aromatic hydrocarbons (PAHs), Benzene.