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An outline of the PCR design for detecting the H3.3K27M mutant allele. The codon for lysine 27 of the H3F3A gene is AAG and is mutated to ATG at one allele of the H3F3A gene. The 3’-end of each mutant-specific primer ends at mutant variant-base (T). The blocker oligo was phosphorylated at its 3’-end to prevent elongation, with the wild type base corresponding to the mutant allele placed in the center of the sequence. A reverse primer is also indicated.
