O6-Methylguanine–DNA Methyltransferase (MGMT) Promoter Methylation Analysis in Glioblastoma Patients

The alkylating agent temozolomide is used to treat patients who have glioblastoma. However, treatment is only effective when O6-methylguanine–DNA methyltransferase is silenced in the cancer cells. The best way to assess whether or not a patient will be responsive to treatment is to determine if the promoter region of the O6-methylguanine–DNA methyltransferase gene is methylated. Similarly, knowing the methylation status of the O6-methylguanine–DNA methyltransferase promoter in patient -derived brain tumor initiating cells can help guide research. Here we describe how to determine the methylation status in vitro using bisulfite conversion followed by methylation-specific PCR.