Approach to somatic mutation discovery. After identifying genes of interest, the Celera and public genome databases were used to extract exon sequences and intronic regions to design primers for PCR amplification and sequencing. Successfully amplified and sequenced exons were assembled and compared with exon reference sequences to identify nonsynonymous alterations. Any potential alterations were then compared with polymorphism databases to exclude known SNPs. For remaining alterations, PCR products from matched normal and tumor DNA were sequenced to identify novel SNPs and somatic mutations.

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