Fig. 3.

Molecular diagnostics of diffuse gliomas. Use of custom pyrosequencing to detect wild-type (A) and R132H mutant (B, C > T) IDH1 on reverse strand pyrograms. Bisulfite pyrosequencing detects CpG methylation within the MGMT promoter as increased C peaks (≥10%) at five indicated sites (C, negative; D, positive). Lack of the C peak highlighted in gold indicates complete bisulfite conversion. Sanger sequencing detects the 124 C > T (C228T) and 146 C > T (C250T) mutations in the TERT promoter. Only C is present at the indicated position in samples with wild-type TERT promoter (E); a T peak at this site (146) indicates mutation (courtesy of Jonathan Galeotti and Karen Weck).