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At NTUH, genomic DNA extracted from tumour specimens was subjected to real-time polymerase chain reaction (PCR), for which primer pairs encompassing BRAF exon 15 were used (5’-TCATAATGCTTGCTCTGATAGGA-3’ and 5’-GGCCAAAAATTTAATCAGTGGA-3’). Then the amplified DNA fragments were purified and direct sequencing was performed using an automated ABI 3730 sequencer (Applied Biosystems, Foster City, CA, USA) [30]. At KCGMH, the BRAF V600E mutation was identified using the Roche BRAF/NRAS Mutation Test (Roche Diagnostics, IN, USA) as per the manufacturer’s instructions.
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