All SNP discovery and genotyping was performed using fluorescent Sanger sequencing of PCR templates generated directly from a subject's genomic DNA. Primers flanking the ALK exons were used to directly isolate sequencing template from genomic DNA. Sequencing/genotyping was performed by fluorescent Sanger chemistry as implemented by Applied Biosystems (Life Technologies) directly applied to the PCR amplified ALK exons. The sequences generated were analyzed for the presence of SNPs and genotyped using the software package Mutation Surveyor from SoftGenetics LLC (State College, PA, USA). To maximize sensitivity and accuracy, all analysis was performed on both strands and all heterozygote calls were verified by human inspection.
