Fig. 3

H3K27M Detection and Validation in Patient CSF and Tumor Tissue Specimens. a CSF-derived DNA and DNA from matched fresh frozen DIPG tumor tissue (PID 2) was submitted for PCR-amplification of a 300 bp region of H3F3A for mutation detection. Sanger sequencing chromatograph of resulting PCR-amplified H3F3A confirmed c.83A > T transversion in CSF DNA and matched DIPG tumor tissue DNA (arrow). b CSF-derived DNA and matched fresh frozen paraffin embedded (FFPE) tumor tissue from PID 10 and 11 was submitted for PCR-amplification of a 300 bp region of H3F3A for mutation detection. Sanger sequencing of resulting PCR-amplified H3F3A from CSF and FFPE tumor tissue demonstrated absence of mutation. c Targeted H3F3A c.83A > T amplification using CSF-derived DNA from PID 1 and 5 demonstrated presence of mutation, with DNA from H3.3K27M DIPG tissue (PID 2) and primary tumor cells (SF8628) as positive controls