DNA methylation in pancreatic cancer: protocols for the isolation of DNA and bisulfite modification.

Pancreatic ductal adenocarcinoma (PDAC) is an aggressive tumor and still remains a challenge for its lack of effective therapeutic strategies, which is due to the late diagnosis of this disease. Methylation markers might improve early detection and surveillance of PDAC. Furthermore, analysis of hypermethylation in the tumor tissue might help to identify new targets for therapeutic intervention and improve the understanding of the pathophysiological changes occurring in pancreatic cancer. Methylation specific PCR is the method of choice if a small number of genes will be tested in a larger set of patient samples. After DNA isolation by standard procedure, the DNA is then modified using sodium bisulfite. This DNA can then be used in qualitative and quantitative PCR assays.